Lack of HLA class I antigen expression by cultured melanoma cells FO-1 due to a defect in B2m gene expression.

C, D'Urso; Wang, Zhigang; Cao, Yan; Tatake, Revati J.; Zeff, Richard A.; Ferrone, Soldano

doi:10.1172/jci114984

Cited by 277 publications

(175 citation statements)

References 58 publications

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“…These heavy chains, 'free' of β 2 m, have been demonstrated to be either 'unfolded' (Allen et al, 1986;Smith et al, 1993), or at least partially folded (Machold et al, 1995), and to bear mature carbohydrates (Hansen et al, 1988;Machold et al, 1995). Biochemical and functional data in β 2 m knockout mice are consistent with the idea that both D b -and L d -free heavy chains may retain some ability to bind (Smith et al, 1993) and present (Lehmann-Grube et al, 1994;Cook et al, 1995) antigen even when synthesized in the absence of β 2 m. In contrast, there is no evidence that any of the more than 300 sequenced HLA-A and -B alleles might be constitutively expressed on the cell surface of most (Fellous et al, 1977;D'Urso et al, 1992;Gattoni-Celli et al, 1992;Wang et al, 1993;Bicknell et al, 1994;Porgador et al, 1997) β 2 m-defective cells of human origin.…”

supporting

confidence: 75%

“…The previously defined HLA-A25 and -B8 specificities (D'Urso et al, 1992) were found to correspond to the HLA-A*2501 and -B*0801 alleles. HLA-C, for which no tissue typing is available, was genotyped as HLA-Cw*0701.…”

Section: Hla-a -B -C Genotyping Of Fo-1 Cellsmentioning

confidence: 70%

“…The β 2 m-defective FO-1 melanoma cell line and its serological HLA typing (A25, -B8) have been previously described (D'Urso et al, 1992). The HLA-A, -B, -C genotype of FO-1 was assessed by locus-specific polymerase chain reaction (PCR) amplification of genomic DNA followed by second and third exon direct sequencing, as described (Pera et al, 1997).…”

Section: Cell Lines and Nucleic Acid Biochemistrymentioning

confidence: 99%

“…In the present report, we have studied the conformation, intracellular transport and expression of class I molecules in the β 2 m-defective FO-1 melanoma cell line (D'Urso et al, 1992) and its β 2 m transfectants. To this end, we have used monoclonal antibodies detecting free and β 2 m-associated HLA-A, -B, -C heavy chains and have taken advantage of a recently characterized (Setini et al, 1996) monoclonal antibody, named L31.…”

mentioning

confidence: 99%

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Biosynthesis of HLA-C heavy chains in melanoma cells with multiple defects in the expression of HLA-A, -B, -C molecules

et al. 1999

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Summary Recent investigations have shown that malignant transformation may down-regulate the expression of class I HLA molecules, β 2 -microglobulin (β 2 m) and members of the antigen-processing machinery. In the present study, we HLA-genotyped and identified at a biochemical level the three (HLA-A25, -B8, -Cw7) class I alleles expressed by the previously described [D'Urso CM et al (1992) J Clin Invest 87: 284-292] β 2 m-defective human melanoma FO-1 cell line and tested their ability to interact with calnexin, calreticulin and the TAP (transporter associated with antigen processing) complex. All these alleles were found to bind calnexin, but not calreticulin or the poorly expressed TAP complex, both in parental and β 2 m-transfected FO-1 cells, demonstrating a complex defect of class I expression in FO-1 cells. In these conditions, Cw7 heavy chains interacted with calnexin more strongly than A25 and B8, and preferentially accumulated in the endoplasmic reticulum, in both a calnexin-associated and a calnexin-free form. In addition, they could be transported to the cell surface at low levels even in the absence of β 2 m, without undergoing terminal glycosylation. These results establish a parallel between HLA-C and the murine D b and L d molecules which have been found to be surface expressed and functional in β 2 m-defective cells. They also demonstrate distinctive features of HLA-C molecules. We propose that the accumulation of several assembly intermediates of HLA-C might favour the binding of peptide antigens not readily bound by HLA-A and -B molecules in neoplastic cells with suboptimal class I expression.

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supporting

confidence: 75%

Section: Hla-a -B -C Genotyping Of Fo-1 Cellsmentioning

confidence: 70%

Section: Cell Lines and Nucleic Acid Biochemistrymentioning

confidence: 99%

mentioning

confidence: 99%

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Biosynthesis of HLA-C heavy chains in melanoma cells with multiple defects in the expression of HLA-A, -B, -C molecules

et al. 1999

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“…Our explanation for this finding could be that heavy chains recognized by HCA2 antibody (mainly of the HLA-A type) might be more stable at the cell surface in the absence of β2-m than the heavy chains recognized by HC10 (mainly of the HLA-B/C types). The underlying mechanism of β2-m weak expression could involve deletion or mutation in the β2-m gene, in analogy with other well studied tumours (Klein et al, 1967;Rosa et al, 1983;D'Urso et al, 1991;Bicknell et al, 1994). Alternatively, it could result from a post-translational mechanism such as sequestering of the β2-m protein by a viral or other protein, in analogy with the mechanism described for cytomegalovirus down-regulation of MHC class I molecules (Chapman and Bjorkman, 1998).…”

Section: Mhc Class I Expression In Osccsmentioning

confidence: 96%

Lack of MHC class I surface expression on neoplastic cells and poor activation of the secretory pathway of cytotoxic cells in oral squamous cell carcinomas

et al. 1999

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Cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells use the secretory pathway of perforin/granzymes to kill their target cells. In contrast to NK cells, CTL responses are MHC class I restricted. In this study we analysed the relative activation of CTL and NK cells in relation with MHC class I expression on oral squamous cell carcinomas (OSCCs). MHC class I expression was investigated in 47 OSCCs by immunohistochemistry using HCA2, HC10 and β2-m antibodies. The presence of CTLs, NK cells, and its activation, was investigated in 21 of these OSCCs using respectively, CD8, CD57 and GrB7 antibodies. The Q-Prodit measuring system was used for quantification of cytotoxic cells. All OSCCs showed weak or absent staining of β2-m on the cell surface. The absence of β2-m was significantly associated with absent expression of MHC class I heavy chain as detected by HC10 antibody ( P = 0.004). In tumour infiltrates CTLs always outnumbered NK cells, as reflected by the ratio CD57/CD8 being always inferior to one (mean: 0.19; SD: 0.15). The proportion of activated cytotoxic cells as detected by granzyme B expression was generally low (mean: 8.6%; SD 8.9). A clear correlation between MHC class I expression and the relative proportion of NK cells/CTLs was not found. This study shows that the majority of OSCCs show weak or absent expression of MHC class I molecules on the cell surface, possibly due to alterations in the normal β2-m pathway. The low proportion of granzyme B-positive CTLs/NK cells indicates that the secretory pathway of cytotoxicity is poor in these patients. The lack of correlation between MHC class I expression and CTL/NK cell activation as detected by granzyme B expression suggests that, next to poor antigen presentation, also local factors seem to determine the final outcome of the cytotoxic immune response. © 1999 Cancer Research Campaign

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Structural and functional analysis of β2 microglobulin abnormalities in human lung and breast cancer

et al. 1996

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The escape of tumor cells from immune recognition is a central problem in tumor immunology. Here, we examined the functional role of somatic beta 2-microglobulin (beta2m) gene mutations in human lung and breast cancers. Using single-strand conformational polymorphism (SSCP) analysis and DNA sequencing, we found mutations in the beta2m gene in 2 of 110 tested lung, colon and breast tumors and tumor cell lines. No mutations were identified in 63 breast cancer tumors, in B-lymphoblastoid cell lines or normal tissues from these or other patients. In these cell lines, beta2m protein was undetectable by Western blot analysis and there was no MHC class I on their cell surface even after treatment with interferon-gamma. Transfection of these tumor cell lines with the beta2m gene, but not addition of purified beta2m protein restored MHC expression without addition of exogenous pepticles, indicating that endogenous beta2m expression is necessary for proper intracellular MHC assembly and stabilization by endogeneous pepticles. Mutation in beta2m caused cell line H2009 to be resistant to specific lysis by influenza virus-specific CTL from HLA matched donors, and transfection of the beta2m gene restored this killing. A small cell lung cancer cell line with low class I expression and with a normal beta2m genomic sequence nonetheless also demonstrated increased class I expression after transfection of the beta2m expression vector alone, indicating that the availability of beta2m may be rate limiting for MHC assembly in this line. Our results indicate that somatic mutations or selective loss of expression of the beta2m gene in human lung cancer is rare, but can cause defective MHC class I expression and function allowing these cells to escape recognition by cytotoxic T cells.

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Lack of HLA class I antigen expression by cultured melanoma cells FO-1 due to a defect in B2m gene expression.

Cited by 277 publications

References 58 publications

Biosynthesis of HLA-C heavy chains in melanoma cells with multiple defects in the expression of HLA-A, -B, -C molecules

Biosynthesis of HLA-C heavy chains in melanoma cells with multiple defects in the expression of HLA-A, -B, -C molecules

Lack of MHC class I surface expression on neoplastic cells and poor activation of the secretory pathway of cytotoxic cells in oral squamous cell carcinomas

Structural and functional analysis of β2 microglobulin abnormalities in human lung and breast cancer

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