HLA-G, -E, -F preworkshop: tools and protocols for analysis of non-classical class I genes transcription and protein expression

Paul, Pascale; Rouas‐Freiss, Nathalie; Moreau, Philippe; Cabestre, Francisco Adrian; Menier, Catherine; Khalil-Daher, Iman; Pangault, Céline; Onno, Myriam; Fauchet, R.; Martı́nez-Laso, Jorge; Morales, Pablo; Villena, Antonio Arnaiz; Giacomini, Patrizio; Natali, Pier Giorgio; Frumento, Guido; Ferrara, G. B.; McMaster, Michael; Fisher, Susan J.; Schust, Danny J.; Ferrone, Soldano; Dausset, J; Geraghty, Dan; Carosella, Edgardo D.

doi:10.1016/s0198-8859(00)00154-3

Cited by 116 publications

(102 citation statements)

References 47 publications

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“…To visualize the alternative mRNA forms, HLA-G transcripts were analyzed by reverse transcription-PCR (RT-PCR) and Southern blot hybridization, using procedures validated at the 13th International HLA Workshop. 34 As shown in Figure 3a, the HLA-G alternative splicing profile of Fon 1 cells is similar to that of JEG-3 cells with a predominance of HLA-G1/G5 transcripts and lower levels of HLA-G2/G4 transcripts. Interestingly, Fon 2 cells exhibited a totally different profile with an absence of HLA-G1 transcript and upregulation of HLA-G2/G4 transcripts.…”

Section: Hla-g2 Mrna and Protein Expression Is Switched On In Hla-g1-supporting

confidence: 53%

“…The M8-pcDNA cells (transfected with the control vector alone) and the M8-HLA-G1, -HLA-G2, -HLA-G3, -HLA-G4 and -HLA-G5 cells (transfected with the vector containing either the HLA-G1, HLA-G2, HLA-G3, HLA-G4 or HLA-G5 cDNA) were obtained as previously described and selected in media containing 100 lg/ml hygromicin B (Sigma Chemical Co.). 23,34 The cells used were routinely tested for and found to be free of mycoplasma.…”

Section: Cell Lines and Antibodiesmentioning

confidence: 99%

“…The following MAbs (monoclonal antibodies) were used: MEM-G/ 9, a mouse IgG1 specific for native HLA-G1 and HLA-G5 isoforms (Exbio, Prague, Czech Republic); 35 4H84, a mouse IgG1 anti-HLA-G free heavy chain (kindly provided by M. Mc Master, University of California, San Francisco, CA); 34,36 5A6G7, a mouse IgG1 specific for the intron 4-retaining part of both soluble HLA-G5 and HLA-G6; 37 TP25.99, a mouse IgG1 anti-HLA-A, -B, -C and -E but not anti-HLA-G (kindly provided by S. Ferrone, Roswell Park Cancer Institute, Buffalo, NY); 34,38 B1G6, a mouse IgG2a, anti-b2 microglobulin (Immunotech, France); B8.12.2, a mouse IgG2b anti-HLA-DR (Immunotech); GHI/75, a mouse IgG2b anti-ILT-2 (Pharmingen, San Diego, CA); Z199, a mouse IgG2b anti-NKG2A (Immunotech) and B-5-1-2 , a mouse IgG1 anti-a tubulin (Sigma Chemical Co.).…”

Section: Cell Lines and Antibodiesmentioning

confidence: 99%

“…Immunostaining was evaluated on tissues using the DAKO EnVision 1 System, Peroxidase (AEC) (Dako, France), as previously described. 34 …”

Section: Immunohistochemistry Analysismentioning

confidence: 99%

“…For HLA-G1 to -G5 transcript detection, PCR amplification and hybridization of the PCR products were performed as previously described. 34,39 The JEG-3 cell line was used as an HLA-G transcription positive control.…”

Section: Isolation Of Rna Reverse Transcription Pcr Amplification Amentioning

confidence: 99%

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Switch ofHLA-G alternative splicing in a melanoma cell line causes loss of HLA-G1 expression and sensitivity to NK lysis

et al. 2005

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these points by studying a melanoma cell line derived from a surgically-removed HLA-G-positive melanoma lesion. We show that HLA-G expression in melanoma cells can be regulated at the mRNA splicing level. Indeed, melanoma cells rapidly switched from cell-surface HLA-G1 to intra-cellular HLA-G2 expression. This mechanism restored tumor sensitivity to NK lysis. Moreover, switch from HLA-G1 to HLA-G2 was strong enough to prevent re-expression of immunoprotective HLA-G1 even following treatments with cytokines and DNA demethylating agent. Modulating HLA-G at the mRNA splicing level would be an efficient way of lifting in vivo HLA-G-mediated tumor immune escape. ' 2005 Wiley-Liss, Inc.

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Section: Hla-g2 Mrna and Protein Expression Is Switched On In Hla-g1-supporting

confidence: 53%

Section: Cell Lines and Antibodiesmentioning

confidence: 99%

Section: Cell Lines and Antibodiesmentioning

confidence: 99%

“…Immunostaining was evaluated on tissues using the DAKO EnVision 1 System, Peroxidase (AEC) (Dako, France), as previously described. 34 …”

Section: Immunohistochemistry Analysismentioning

confidence: 99%

Section: Isolation Of Rna Reverse Transcription Pcr Amplification Amentioning

confidence: 99%

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Switch ofHLA-G alternative splicing in a melanoma cell line causes loss of HLA-G1 expression and sensitivity to NK lysis

et al. 2005

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Novel landscape of HLA‐G isoforms expressed in clear cell renal cell carcinoma patients

et al. 2017

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Immune checkpoints are powerful inhibitory molecules that promote tumor survival. Their blockade is now recognized as providing effective therapeutic benefit against cancer. Human leukocyte antigen G (HLA‐G), a recently identified immune checkpoint, has been detected in many types of primary tumors and metastases, in malignant effusions as well as on tumor‐infiltrating cells, particularly in patients with clear cell renal cell carcinoma (ccRCC). Here, in order to define a possible anticancer therapy, we used a molecular approach based on an unbiased strategy that combines transcriptome determination and immunohistochemical labeling, to analyze in‐depth the HLA‐G isoforms expressed in these tumors. We found that the expression of HLA‐G is highly variable among tumors and distinct areas of the same tumor, testifying a marked inter‐ and intratumor heterogeneity. Moreover, our results generate an inventory of novel HLA‐G isoforms which includes spliced forms that have an extended 5′‐region and lack the transmembrane and alpha‐1 domains. So far, these isoforms could not be detected by any method available and their assessment may improve the procedure by which tumors are analyzed. Collectively, our approach provides the first extensive portrait of HLA‐G in ccRCC and reveals data that should prove suitable for the tailoring of future clinical applications.

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Increased telomerase activity is associated with shorter survival in patients with chronic phase chronic myeloid leukemia

Verstovšek

Kantarjian

Manshouri

et al. 2003

Cancer

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HLA-G, -E, -F preworkshop: tools and protocols for analysis of non-classical class I genes transcription and protein expression

Cited by 116 publications

References 47 publications

Switch ofHLA-G alternative splicing in a melanoma cell line causes loss of HLA-G1 expression and sensitivity to NK lysis

Switch ofHLA-G alternative splicing in a melanoma cell line causes loss of HLA-G1 expression and sensitivity to NK lysis

Novel landscape of HLA‐G isoforms expressed in clear cell renal cell carcinoma patients

Increased telomerase activity is associated with shorter survival in patients with chronic phase chronic myeloid leukemia

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