Identification of CD19 and CD20 Peptides for Induction of Antigen-Specific CTLs against B-Cell Malignancies

Bae, Jooeun; Martinson, Jeffrey; Klingemann, Hans

doi:10.1158/1078-0432.ccr-04-1612

Cited by 21 publications

(20 citation statements)

References 51 publications

(26 reference statements)

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“…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%

“…To evaluate the presence of D393-CD20 specific T cell in the human repertoire, the D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] 20mer-peptide overlapping the splicing site was used to stimulate T cells. For this purpose, peripheral blood lymphocytes of healthy volunteers were in vitro stimulated twice using 10 mmol/L of D393-CD20 [28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47] in 96 well microplates during 14 days as described in the material and method section.…”

Section: D393-cd20 Isoform Protein Expression and Immunogenicitymentioning

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: Mouse and Vaccinationsmentioning

confidence: 99%

Section: D393-cd20 Isoform Protein Expression and Immunogenicitymentioning

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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Invariant chain as a vehicle to load antigenic peptides on human MHC class I for cytotoxic T‐cell activation

et al. 2013

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IntroductionSuccessful vaccination depends on activation of CD8 + and CD4 + T-cell responses by presentation of antigenic peptides in the context of major histocompatibility complex class I (MHCI) and II Correspondence: Dr. Tone F. Gregers e-mail: t.f.gregers@ibv.uio.no (MHCII) molecules, respectively. The processing and subsequent presentation of antigenic peptides by MHCI or MHCII molecules on the surface of antigen presenting cells (APCs) requires a coordinated action of different accessory molecules and dedicated chaperones. Targeting of Ags to these pathways that * These authors share senior co-authorship.C 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu Eur. J. Immunol. 2014. 44: 774-784 Immunomodulation 775 improve presentation to T cells, would therefore be expected to potentiate vaccine strategies. MHCII interacts with the dedicated chaperone invariant chain (Ii) in the ER. Ii contains sorting signals within its cytoplasmic tail, which mediate trafficking of Ii-MHCII to proteolytic endosomal compartments where Ag loading takes place [1]. In the endosomal pathway Ii is sequentially degraded, leaving a residual CLIP in the peptide-binding groove of MHCII, which is exchanged by specific antigenic peptides by the action of HLA-DM.Indeed, Ags targeted to the endosomal pathway for loading onto MHCII have successfully been shown to activate CD4 + T-cell responses. Ag targeting has been achieved using signals associated with lysosomal proteins, such as lysosomal-associated membrane protein 1 [2] or other lysosomal hydrolases [3], by direct coupling of the Ag to full-length Ii, or to Ii containing the endosomal localization signal only [4,5]. Finally, genetic exchange of the CLIP sequence with antigenic epitopes was demonstrated to be highly efficient for loading of MHCII and CD4 + T-cell activation [6]. In fact, this strategy has proven to be superior compared to peptide loading for priming of CD4 + T cells and tumor protection in vivo [7, 8].Loading of MHCI molecules for vaccine purposes has commonly been achieved by exogenous pulsing of APCs with synthetic peptides in vitro. This approach is limited by rapid degradation of peptides, the risk of inducing T-cell tolerance [9], and the possibility for the formation of cryptic epitopes leading to imprecise tumor targeting [10]. Another alternative is Ag delivery by mRNA [11] or DNA [12], relying on protein production and proteasomal degradation to generate high peptide concentrations in the cytosol of the APC. Peptides are subsequently translocated into the ER by transporters associated with Ag processing (TAP), and loaded onto MHCI [11]. However, vaccination strategies that are independent of proteasomal degradation might represent an advantage in several contexts. Some tumor Ags are processed more efficiently by the immunoproteasomes of dendritic cells (DCs) than by the standard proteasomes present in the majority of other cells, whereas for others the situation is vice versa [13,14]. Moreover, the combination of immunotherapy with certain...

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A novel interaction of nucleophosmin with BCL2-associated X protein regulating death evasion and drug sensitivity in human hepatoma cells

Fan

Tsai

et al. 2013

Hepatology

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Death evasion is crucial for both carcinogenesis and resistance to anticancer therapies. Recently, we identified nucleophosmin (NPM) as a key factor counteracting death stimuli in human hepatocellular carcinoma (HCC) cells. Here we report the identification of a novel NPM-BCL2-associated X protein (BAX) pathway orchestrating death evasion in human HCC cells. Silencing of NPM expression significantly sensitized HCC cells-particularly those bearing inactivated p53 gene (Huh7, Hep3B, and Mahlavu)-to ultraviolet irradiation, mitomycin C, doxorubicin, cisplatin, sorafenib, and lapatinib. This sensitizing effect was not changed further, as p53 expression had been simultaneously silenced. Following cell stress, NPM and BAX were induced and exported out of the nucleoli and nucleus, respectively. BAX was translocated to cytoplasm in cells with relatively high NPM level, or accumulated in the mitochondria in cells with relatively low NPM level and undergoing apoptosis. Subcellular fractionation revealed that silencing of NPM expression greatly enhanced mitochondrial translocation and oligomerization of BAX in Huh7 and Mahlavu cells. In situ proximity ligation assays and reciprocal co-immunoprecipitation revealed a direct interaction between NPM and BAX in the cytoplasm. Silencing of BAX expression abolished the sensitization effect exerted by silencing of NPM in HCC cells. Clinically, up-regulation of NPM was significantly associated with advanced tumor stage and poor prognosis. Conclusion: By directly blockading BAX mitochondrial translocation and activation, NPM helps human HCC cells evade death induction independently of p53-mediated cell death. Silencing of NPM significantly sensitized HCC cells to anticancer therapies. NPM is a potential cotarget in combination with other therapies for HCC, particularly those that harbor inactivated p53 gene. Our findings are of clinical significance because NPM up-regulation and p53 mutations are usually found in advanced human cancers, including HCC.

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Identification of CD19 and CD20 Peptides for Induction of Antigen-Specific CTLs against B-Cell Malignancies

Cited by 21 publications

References 51 publications

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

CD20 alternative splicing isoform generates immunogenic CD4 helper T epitopes

Invariant chain as a vehicle to load antigenic peptides on human MHC class I for cytotoxic T‐cell activation

A novel interaction of nucleophosmin with BCL2-associated X protein regulating death evasion and drug sensitivity in human hepatoma cells

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