Abstract
We developed a novel human leukocyte antigen HLA–ABC locus-specific quantitative real-time polymerase chain reaction (PCR) to determine the locus-specific gene expression of HLA–ABC in peripheral blood leukocytes (PBLs, n = 53), colon mucosa (n = 15), and larynx mucosa (n = 15). Laser-assisted tissue microdissection allowed us to study the selected cells without interference from surrounding stroma. We report evidence on the specificity of the technique, describing the HLA–ABC locus-specific gene expression patterns found in the PBLs and two solid tissues studied. PBLs showed a higher gene expression of HLA-B than of HLA-A or HLA-C (p = 4.7 × 10−10 and p = 1.6 × 10−6, respectively). In solid tissue, HLA-A and HLA-B gene expressions were similar and HLA-C expression lower. In particular, in larynx mucosa, significant differences were found between HLA-A and HLA-C expressions and between HLA-B and HLA-C expressions (p = 6.5 × 10−4 and p = 8.1 × 10−4, respectively). The same differences were observed in colon mucosa, but significance was not reached (p = 0.08 and p = 0.06, respectively). Differences in locus-specific regulation may be related to the control of cytotoxic responses of NK and CD8 positive T cells. Gene expression of HLA–ABC specific locus showed no intra-individual variability, but there was a high inter-individual variability. This may result from differences in the expression of common regulatory factors that control HLA–ABC constitutive expression.
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References
Aptsiauri N, Cabrera T, Garcia-Lora A, Lopez-Nevot MA, Ruiz-Cabello F, Garrido F (2007) MHC class I antigens and immune surveillance in transformed cells. Int Rev Cytol 256:139–189
Burrone OR, Milstein C (1982) Control of HLA-A, B, C synthesis and expression in interferon-treated cells. EMBO J 1:345–349
Cabrera T, Lopez-Nevot MA, Gaforio JJ, Ruiz-Cabello F, Garrido F (2003) Analysis of HLA expression in human tumor tissues. Canc Immunol Immunother 52:1–9
Campoli M, Chang CC, Ferrone S (2002) HLA class I antigen loss, tumor immune escape and immune selection. Vaccine 20(Suppl 4):A40–A45
Collins KL, Baltimore D (1999) HIV's evasion of the cellular immune response. Immunol Rev 168:65–74
Daar AS, Fuggle SV, Fabre JW, Ting A, Morris PJ (1984) The detailed distribution of HLA-A, B, C antigens in normal human organs. Transplantation 38:287–292
Garrido F, Algarra I (2001) MHC antigens and tumor escape from immune surveillance. Adv Cancer Res 83:117–158
Garrido F, Ruiz-Cabello F, Cabrera T, Perez-Villar JJ, Lopez-Botet M, Duggan-Keen M, Stern PL (1997) Implications for immunosurveillance of altered HLA class I phenotypes in human tumours. Immunol Today 18:89–95
Johnson DR (2000) Differential expression of human major histocompatibility class I loci: HLA-A, -B, and -C. Hum Immunol 61:389–396
Le Morvan C, Cogne M, Drouet M (2001) HLA-A and HLA-B transcription decrease with ageing in peripheral blood leucocytes. Clin Exp Immunol 125:245–250
Liu K, Kao KJ (1997) Measurement of relative quantities of different HLA-A and -B mRNAs in cells by reverse transcription-polymerase chain reaction and denaturing gradient gel electrophoresis. J Immunol Meth 203:67–75
Liu K, Kao KJ (2000) Mechanisms for genetically predetermined differential quantitative expression of HLA-A and -B antigens. Hum Immunol 61:799–807
Majumder D, Bandyopadhyay D, Chandra S, Mukhopadhayay A, Mukherjee N, Bandyopadhyay SK, Banerjee S (2005) Analysis of HLA class Ia transcripts in human leukaemias. Immunogenetics 57:579–589
Maleno I, Cabrera CM, Cabrera T, Paco L, Lopez-Nevot MA, Collado A, Ferron A, Garrido F (2004) Distribution of HLA class I altered phenotypes in colorectal carcinomas: high frequency of HLA haplotype loss associated with loss of heterozygosity in chromosome region 6p21. Immunogenetics 56:244–253
Mendez R, Rodriguez T, Del Campo A, Monge E, Maleno I, Aptsiauri N, Jimenez P, Pedrinaci S, Pawelec G, Ruiz-Cabello F, Garrido F (2008) Characterization of HLA class I altered phenotypes in a panel of human melanoma cell lines. Cancer Immunol Immunother 57:719–729
Middleton D, Curran M, Maxwell L (2002) Natural killer cells and their receptors. Transpl Immunol 10:147–164
Myers NB, Wormstall E, Hansen TH (1996) Differences among various class I molecules in competition for beta2m in vivo. Immunogenetics 43:384–387
Paco L, Garcia-Lora AM, Casares C, Cabrera C, Algarra I, Collado A, Maleno I, Garrido F, Lopez-Nevot MA (2007) Total loss of HLA class I expression on a melanoma cell line after growth in nude mice in absence of autologous antitumor immune response. Int J Cancer 121:2023–2030
Robinson J, Marsh SG (2007) The IMGT/HLA database. Meth Mol Biol 409:43–60
Romero JM, Jimenez P, Cabrera T, Cozar JM, Pedrinaci S, Tallada M, Garrido F, Ruiz-Cabello F (2005) Coordinated downregulation of the antigen presentation machinery and HLA class I/beta2-microglobulin complex is responsible for HLA–ABC loss in bladder cancer. Int J Cancer 113:605–610
Serrano A, Tanzarella S, Lionello I, Mendez R, Traversari C, Ruiz-Cabello F, Garrido F (2001) Rexpression of HLA class I antigens and restoration of antigen-specific CTL response in melanoma cells following 5-aza-2′-deoxycytidine treatment. Int J Cancer 94:243–251
Shieh DC, Gammon MC, Zweerink HJ, Kao KJ (1996) Functional significance of varied quantitative and qualitative expression of HLA-A2.1 antigens in determining the susceptibility of cells to cytotoxic T lymphocytes. Hum Immunol 46:18–26
Tanzarella S, Russo V, Lionello I, Dalerba P, Rigatti D, Bordignon C, Traversari C (1999) Identification of a promiscuous T-cell epitope encoded by multiple members of the MAGE family. Canc Res 59:2668–2674
Thomas R, Apps R, Qi Y, Gao X, Male V, O'hUigin C, O'Connor G, Ge D, Fellay J, Martin JN, Margolick J, Goedert JJ, Buchbinder S, Kirk GD, Martin MP, Telenti A, Deeks SG, Walker BD, Goldstein D, McVicar DW, Moffett A, Carrington M (2009) HLA-C cell surface expression and control of HIV/AIDS correlate with a variant upstream of HLA-C. Nat Genet 41:1290–1294
Trowsdale J, Betz AG (2006) Mother's little helpers: mechanisms of maternal-fetal tolerance. Nat Immunol 7:241–246
van den Elsen PJ, Holling TM, Kuipers HF, van der Stoep N (2004) Transcriptional regulation of antigen presentation. Curr Opin Immunol 16:67–75
Versteeg R, Kruse-Wolters KM, Plomp AC, van Leeuwen A, Stam NJ, Ploegh HL, Ruiter DJ, Schrier PI (1989) Suppression of class I human histocompatibility leukocyte antigen by c-myc is locus specific. J Exp Med 170:621–635
Acknowledgments
The authors thank Ana Isabel Rodríguez Fernández for technical assistance. The study was partially supported by grants from the Fondo de Investigaciones Sanitarias (08/1265), Red Genómica del Cáncer (RETICRD 06/020), Consejería de Salud Junta de Andalucía, Proyecto de Investigación de Excelencia (CTS-3952 and CVI-4740) and Plan Andaluz de Investigación (PAI, Group CTS-143) in Spain, and from the European Searchable Tumour Cell Line Database (ESTDAB) project, contract No. QLRI-CT-2001-01325 (http://www.ebi.ac.uk/estdab), from the European Network for the identification and validation of antigens and biomarkers in cancer and their application in clinical tumor immunology (ENACT) project (European community LSHC-CT-2004-503306) and from the Cancer Immunotherapy project (European community OJ 2004/c158,18234)
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García-Ruano, A.B., Méndez, R., Romero, J.M. et al. Analysis of HLA–ABC locus-specific transcription in normal tissues. Immunogenetics 62, 711–719 (2010). https://doi.org/10.1007/s00251-010-0470-z
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DOI: https://doi.org/10.1007/s00251-010-0470-z