A comparison of the immunogenicity of the native and denatured forms of a protein

Koch, Claus; Jensen, Sanne; Øster, Anne

doi:10.1111/j.1699-0463.1996.tb00696.x

Cited by 37 publications

(31 citation statements)

References 27 publications

(5 reference statements)

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“…Although the fractionation process is carried out under partially denaturing conditions, antigen uptake, processing, and cross-presentation by DCs are not impaired. This is in accordance with previous observations that protein denaturation increases uptake, processing, and presentation by DCs and thus increases overall immunogenicity (26)(27)(28). Importantly, the amounts of purified proteins obtained from the separation process were sufficient for recognition by memory T cells.…”

Section: Discussionsupporting

confidence: 92%

“…immunogenic epitope in more detail, we synthesized overlapping 20-meric peptides of the respective region ( Figure 6C) and identified S100A9 [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] as the immunogenic epitope ( Figure 6D). This epitope was recognized both by CD8 + and CD4 + T cells of patient NCH550 ( Figure 6E).…”

Section: Figurementioning

confidence: 99%

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Rapid T cell–based identification of human tumor tissue antigens by automated two-dimensional protein fractionation

Beckhove¹,

Warta²,

Lemke³

et al. 2010

J. Clin. Invest.

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Identifying the antigens that have the potential to trigger endogenous antitumor responses in an individualcancer patient is likely to enhance the efficacy of cancer immunotherapy, but current methodologies do not efficiently identify such antigens. This study describes what we believe to be a new method of comprehensively identifying candidate tissue antigens that spontaneously cause T cell responses in disease situations. We used the newly developed automated, two-dimensional chromatography system PF2D to fractionate the proteome of human tumor tissues and tested protein fractions for recognition by preexisting tumor-specific CD4 + Th cells and CTLs. Applying this method using mice transgenic for a TCR that recognizes an OVA peptide presented by MHC class I, we demonstrated efficient separation, processing, and cross-presentation to CD8 + T cells by DCs of OVA expressed by the OVA-transfected mouse lymphoma RMA-OVA. Applying this method to human tumor tissues, we identified MUC1 and EGFR as tumor-associated antigens selectively recognized by T cells in patients with head and neck cancer. Finally, in an exemplary patient with a malignant brain tumor, we detected CD4 + and CD8 + T cell responses against two novel antigens, transthyretin and calgranulin B/S100A9, which were expressed in tumor and endothelial cells. The immunogenicity of these antigens was confirmed in 4 of 10 other brain tumor patients. This fast and inexpensive method therefore appears suitable for identifying candidate T cell antigens in various disease situations, such as autoimmune and malignant diseases, without being restricted to expression by a certain cell type or HLA allele.

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Section: Discussionsupporting

confidence: 92%

Section: Figurementioning

confidence: 99%

Rapid T cell–based identification of human tumor tissue antigens by automated two-dimensional protein fractionation

Beckhove¹,

Warta²,

Lemke³

et al. 2010

J. Clin. Invest.

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“…4e). Because the two anti-OVA mAbs are reported to recognize different nonoverlapping epitopes on native OVA protein (38) and do not react with SIINFEKL peptide, these results rule out the possibility that small peptides or peptides associated with HSP are the source of antigen for cross-priming. Because the depletion of OVA from the cytosol eliminates cross-priming activity but not hsp70, hsp90, and grp94 (Fig.…”

Section: Resultsmentioning

confidence: 90%

Cellular protein is the source of cross-priming antigenin vivo

Shen

Rock

2004

Proc. Natl. Acad. Sci. U.S.A.

176

129

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Cross-priming is essential for generating cytotoxic T lymphocytes to viral, tumor, and tissue antigens that are expressed exclusively in parenchymal cells. In this process, the antigen-bearing parenchymal cells must somehow transfer their antigens to bone marrow-derived professional antigen-presenting cells. Although intact proteins, small peptides, or peptide-heat shock protein complexes can all be acquired and presented by antigen-presenting cells, the physiologically relevant form of antigen that is actually transferred from parenchymal cells and cross-presented in vivo is unknown and controversial. To address this issue we have investigated the ability of fibroblasts stably expressing chicken ovalbumin constructs targeted to different subcellular compartments to crossprime cytotoxic T lymphocytes. Although these transfectants generated similar amounts of the immunogenic ovalbumin peptide, their cross-priming activity differed markedly. Instead, the cells cross-priming ability correlated with their steady-state levels of ovalbumin protein and͞or the physical form͞location of the protein. Moreover, in subcellular fractionation experiments, the crosspriming activity colocalized with antigenic protein. In addition, depletion of intact protein antigen from these cell fractions eliminated their cross-priming activity. In contrast, the major heat shock protein candidates for cross-presentation were separable from the cell's main sources of cross-priming antigen. Therefore, cellular proteins, rather than peptides or heat shock protein͞ peptide complexes, are the major source of antigen that is transferred from antigen-bearing cells and cross-presented in vivo.

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“…Dietary proteins are usually prepared in heat-denatured and aggregated forms. Owing to different heat treatments, there are physico-chemical discrepancies in the structure of heated proteins [6,7]. Because of the reports that boiled and evaporated cow's milk have a much decreased oral-sensitizing capacity in guinea-pigs [8,9], heat treatment has been recognized as a simple way of reducing allergenicity [9,10].…”

Section: Introductionmentioning

confidence: 99%

Heat Denaturation of Egg‐White Proteins Abrogates the Induction of Oral Tolerance of Specific Th2 Immune Responses in Mice

Peng

Chang²,

Цай

et al. 1998

Scand J Immunol

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Human foods are usually prepared by cooking. Boiling of chicken egg-white (EW) led to decreased allergenicity, and abrogated intestinal uptake of immunoreactive ovalbumin (OVA) when fed to mice. Therefore, the effects of oral administration of boiled EW were examined further in BALB/c mice. Specific IgE, IgG 1 and IgG antibody responses were suppressed by raw EW, but not by EW boiled for 5 or 60 min, fed prior to sensitization with 10 mg OVA or 1 mg DNP-OVA in alum. Similar results were obtained when mice were sensitized with 10 mg conalbumin, ovomucoid or lysozyme in alum. BALB/c spleen cell proliferation and secretion of Th2 cytokines IL-4 and IL-5 during in vitro stimulation with OVA were also suppressed by feeding raw EW, but not by boiled EW. Although heat denaturation of proteins can minimize allergenicity, the present results suggest that over-cooking of proteins may affect their intestinal antigen processing and thus prevent the induction of oral tolerance.

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A comparison of the immunogenicity of the native and denatured forms of a protein

Cited by 37 publications

References 27 publications

Rapid T cell–based identification of human tumor tissue antigens by automated two-dimensional protein fractionation

Rapid T cell–based identification of human tumor tissue antigens by automated two-dimensional protein fractionation

Cellular protein is the source of cross-priming antigenin vivo

Heat Denaturation of Egg‐White Proteins Abrogates the Induction of Oral Tolerance of Specific Th2 Immune Responses in Mice

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