A BCR-ABL Oncoprotein p210b2a2 Fusion Region Sequence Is Recognized by HLA-DR2a Restricted Cytotoxic T Lymphocytes and Presented by HLA-DR Matched Cells Transfected With an Iib2a2 Construct

Blood ◽  
1999 ◽  
Vol 94 (3) ◽  
pp. 1038-1045 ◽  
Author(s):  
George J.A. ten Bosch ◽  
Jan H. Kessler ◽  
Antonia M. Joosten ◽  
Alexandra A. Bres-Vloemans ◽  
Annemieke Geluk ◽  
...  
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
T Cell Responses ◽  
Myelogenous Leukemia ◽  
Invariant Chain ◽  
Specific Cell ◽  
Double Positive ◽  
Cell Responses ◽  
Human T Cell ◽  
Hla Dr

Peptides corresponding to the fusion site in 210 kD BCR-ABL protein b3a2 (p210b3a2) were previously shown to bind to several HLA class I and II alleles. We have found that b3a2 peptide-specific CD4-positive T-helper cells were able to recognize p210b3a2-positive chronic myelogenous leukemia (CML) blasts in a DR4 restricted manner. Until now, there were no reports of b2a2 breakpoint-specific human T-cell responses. Here we show that repetitive stimulation of T lymphocytes with a 17mer peptide covering the fusion region in p210b2a2 also leads to specific T-cell responses. CD4 and CD4/CD8 double-positive clones obtained from a b2a2 peptide-specific cell line were cytotoxic and proliferative in an HLA-DR2a (DRB5*0101) restricted fashion. Autologous Epstein-Barr virus (EBV) transformed cells, expressing BCR-ABLb2a2 on transfection, and allogeneic HLA-DR matched p210b2a2-positive cells from CML patients were, however, not lysed. BCR-ABL peptide-specific T-cell clones did respond to autologous EBV cells transfected with invariant chain (li) cDNA in which the HLA class II–associated invariant chain peptide (CLIP) was replaced by a BCR-ABL b2a2 fusion oligonucleotide sequence, illustrating the potential of these T cells to recognize an endogenous BCR-ABLb2a2ligand.

Superior Acute Myeloid Leukemia-Specific T Cell Responses Using Dendritic Cells Pulsed with Apoptotic Bodies, vs.Tumor Lysates or mRNA.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 295-295
Author(s):  
Dongxia Xing ◽  
William K. Decker ◽  
Sufang Li ◽  
Richard E. Champlin ◽  
John D. MacMannis ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
T Cell ◽  
T Lymphocytes ◽  
T Cell Responses ◽  
Myelogenous Leukemia ◽  
Apoptotic Bodies ◽  
Apoptotic Body ◽  
Tumor Lysate ◽  
Antigen Uptake ◽  
Cell Responses

Abstract Dendritic cells (DCs), which hold promise for cancer immunotherapy have two distinct functions. Immature DCs internalize antigens; while mature DCs trigger native T cell activation. To date, no consensus has been reached concerning the optimal source of tumor antigen nor the mode of antigen uptake for efficient cross-priming of tumor specific response. Reports have demonstrated that human DCs can acquire relevant antigens and stimulate MHC class I-restricted cytotoxic T lymphocytes (CTLs) by phagocytosing apoptotic tumor cells. Apoptosis was established to be the critical trigger of cross-priming in this process. Apoptotic bodies form when cells undergo programmed cell death. In this study, three acute myelogenous leukemia in (AML) antigen preparations were compared in an autologous setting. Apoptotic bodies, Tumor lysate Tumor mRNA Methods: DCs were generated by CD14 magnetic selection of peripheral blood progenitor cells (PBPCs) and six days of culture in GM-CSF and IL-4. Tumor lysate was made by repeated freezing and thawing of patient AML cells (leukodepletion product >95% blasts); mRNA was extracted using a kit (Qiagen); apoptotic bodies were generated with UVB irradiation of a fraction of the same AML product. The rate of apoptosis was detected using Annexin V FITC flow cytometry. The DCs were loaded with mRNA by electroporation. The DCs were loaded with lysate and apoptotic bodies by incubation. Autologous T cells stimulated with DCs loaded with the three differing antigen preparations were compared for interferon(INF)-g production in ELISPOT and proliferation assays. Results: The DCs pulsed with apoptotic bodies; lysate and mRNA expressed similar levels of the costimulatory molecules CD80, CD86, and theDC maturation marker CD83. The internalization rate of apoptotic bodies was about 20% detected by DAPI (label the apoptotic bodies) and DC83 double staining. The lysate pulsed DC and mRNA pulsed DCs were above 90% detected by flow of trace FITC- albumin and tracer GFP mRNA from previous experiments. Yet IFN-g secretion of T lymphocytes was statistically significant higher (P<0.05), using apoptotic body-loaded DC as compared to total lysate or mRNA. The T cell proliferation stimulated by apoptotic body-loaded DC was also better than that of lysate or mRNA loaded DCs (p<0.05). Conclusions: These preliminary data demonstrate that apoptotic body-loaded DCs induced better Th1 T cell responses than DCs loaded with AML lysate or mRNA. One potential advantage conferred by the use of apoptotic body loaded DCs is that presentation of intracellular proteins in tumor lysates and mRNA that are irrelevant to the induction of anti-tumor responses are eliminated, which may lead to better antigen presentation and consequently an improved Th1 response.

scholarly journals An optimized workflow for CRISPR-Cas9 deletion of surface and intracellular factors in primary human T lymphocytes

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247232
Author(s):  
Cristina Leoni ◽  
Niccolò Bianchi ◽  
Lucia Vincenzetti ◽  
Silvia Monticelli
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
Gene Editing ◽  
Protein A ◽  
Surface Protein ◽  
T Cell Responses ◽  
Cell Responses ◽  
Human T Lymphocytes ◽  
Human T Cell ◽  
Non Coding Rna

The appropriate regulation of T lymphocyte functions is key to achieve protective immune responses, while at the same time limiting the risks of tissue damage and chronic inflammation. Deciphering the mechanisms underpinning T cell responses in humans may therefore be beneficial for a range of infectious and chronic diseases. Recently, the development of methods based on CRISPR-Cas9 gene-editing has greatly expanded the available tool-box for the mechanistic studies of primary human T cell responses. While the deletion of a surface protein has become a relatively straightforward task, as long as an antibody for detection is available, the identification and selection of cells lacking an intracellular protein, a non-coding RNA or a protein for which no antibody is available, remain more problematic. Here, we discuss the options currently available to scientists interested in performing gene-editing in primary human T lymphocytes and we describe the optimization of a workflow for the screening and analysis of lymphocytes following gene-editing with CRISPR-Cas9 based on T cell cloning and T7 endonuclease I cleavage assay.

scholarly journals T cell responses to a mixture of Mycobacterium tuberculosis peptides with complementary HLA-DR binding profiles

1996 ◽  
Vol 105 (3) ◽  
pp. 416-421 ◽  
Author(s):  
S. JURCEVIC ◽  
A. HILLS ◽  
G. PASVOL ◽  
R. N. DAVIDSON ◽  
J. IVANYI ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
T Cell ◽  
T Cell Responses ◽  
Cell Responses ◽  
Hla Dr

Human T-cell responses to oral streptococci in human PBMC-NOD/SCID mice

2006 ◽  
Vol 21 (3) ◽  
pp. 169-176 ◽  
Author(s):  
M. A. Salam ◽  
R. Nakao ◽  
H. Yonezawa ◽  
H Watanabe ◽  
H. Senpuku
Keyword(s):  
T Cell ◽  
T Cell Responses ◽  
Scid Mice ◽  
Oral Streptococci ◽  
Human Pbmc ◽  
Cell Responses ◽  
Human T Cell

scholarly journals Human T cell responses to HPV 16 E2 generated with monocyte-derived dendritic cells

2001 ◽  
Vol 94 (6) ◽  
pp. 807-812 ◽  
Author(s):  
Emma J. Davidson ◽  
Michael D. Brown ◽  
Deborah J. Burt ◽  
Joanna L. Parish ◽  
Kevin Gaston ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
T Cell ◽  
T Cell Responses ◽  
Hpv 16 ◽  
Cell Responses ◽  
Human T Cell

Human T cell responses to the antigen ESAT-6 characterize vaccine candidate and potential diagnostic test for tuberculosis

1999 ◽  
Vol 39 (1) ◽  
pp. A9
Author(s):  
A. Lalvani ◽  
A. Pathan ◽  
R. Brookes ◽  
H. Pritchard ◽  
R. Wilkinson ◽  
...  
Keyword(s):  
T Cell ◽  
Diagnostic Test ◽  
Vaccine Candidate ◽  
T Cell Responses ◽  
Cell Responses ◽  
Human T Cell

scholarly journals BothCD4+andCD8+Lymphocytes Participate in the IFN-γ Response to Filamentous Hemagglutinin fromBordetella pertussisin Infants, Children, and Adults

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Violette Dirix ◽  
Virginie Verscheure ◽  
Françoise Vermeulen ◽  
Iris De Schutter ◽  
Tessa Goetghebuer ◽  
...  
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
Bacterial Infections ◽  
T Cell Responses ◽  
Cell Types ◽  
Production Flow ◽  
Mhc I ◽  
Cell Responses ◽  
Filamentous Hemagglutinin ◽  
Circulating Lymphocytes

Infant CD4+T-cell responses to bacterial infections or vaccines have been extensively studied, whereas studies on CD8+T-cell responses focused mainly on viral and intracellular parasite infections. Here we investigated CD8+T-cell responses uponBordetella pertussisinfection in infants, children, and adults and pertussis vaccination in infants. Filamentous hemagglutinin-specific IFN-γsecretion by circulating lymphocytes was blocked by anti-MHC-I or -MHC-II antibodies, suggesting that CD4+and CD8+T lymphocytes are involved in IFN-γproduction. Flow cytometry analyses confirmed that both cell types synthesized antigen-specific IFN-γ, although CD4+lymphocytes were the major source of this cytokine. IFN-γsynthesis by CD8+cells was CD4+T cell dependent, as evidenced by selective depletion experiments. Furthermore, IFN-γsynthesis by CD4+cells was sometimes inhibited by CD8+lymphocytes, suggesting the presence of CD8+regulatory T cells. The role of this dual IFN-γsecretion by CD4+and CD8+T lymphocytes in pertussis remains to be investigated.

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