868 PRELP-facilitated enhancement of MHC class I surface expression in B16F10 melanoma cells

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A920-A920
Author(s):  
Helene Schäfer ◽  
Karthikeyan Subbarayan ◽  
Barbara Seliger
Keyword(s):  
Cell Proliferation ◽  
Mhc Class I ◽  
Surface Antigens ◽  
Surface Expression ◽  
Human Melanoma ◽  
Class I ◽  
Luciferase Reporter ◽  
List Type ◽  
Promoter Assay ◽  
B16f10 Cells

BackgroundPRELP (proline arginine-rich end leucine-rich repeat protein; also Prolargin), a small leucine-rich proteoglycan, functions as a molecule anchoring basement membranes to connective tissues via the interaction with collagens and heparin. PRELP facilitates the binding of cells to glycosaminoglycans as an important regulator of cell adhesion and thus displays pathophysiological features. Melanoma is an immunogenic tumor, whose relationship with immune cells resident in the microenvironment significantly influences cancer cell proliferation, progression and metastasis. Evasion from immune surveillance is a hallmark of melanoma progression. While our laboratory reported that the proteoglycan biglycan (BGN) was enhancing MHC class I in tumor cells,1 the role of PRELP in tumor immunology has not been studied.MethodsThe murine metastatic melanoma cell line B16F10, characterized by a reduced expression of MHC class I surface antigens was chosen for this study. B16F10 cells were transiently transfected with PRELP as well as co-transfected with BGN. Expression of antigen processing machinery (APM) components and PRELP was determined by qPCR and MHC class I surface expression by flow cytometry. Promoter activity of APM components was analysed by luciferase reporter assays. XTT assays were used to determine cell proliferation. The association of PRELP and MHC class I was studied by bioinformatics in a mixed melanoma dataset of 83 samples.2ResultsOver-expression of PRELP in B16F10 cells enhanced the expression of MHC class I surface antigens, which was due to a PRELP-mediated transcriptional upregulation of components of the MHC class I APM components TAP1, TAP2 and TAPBP as determined by qPCR and promoter assay in PRELP transfectants versus mock controls. Furthermore, MHC class I surface expression was even more pronounced upon BGN co-transfection with PRELP. PRELP overexpression was able to inhibit the proliferation of the B16F10 cells. Bioinformatics analyses demonstrated a positive correlation of PRELP with HLA-A , -B and -C alleles in human melanoma.ConclusionsOur findings demonstrated that overexpression of PRELP correlates with higher MHC class I expression and inhibits cell proliferation. For the first time, co-transfections of the two proteoglycans PRELP and BGN had a synergistic effect on upregulating MHC class I expression. Therefore, PRELP can serve as a novel therapeutic strategy that deserves further investigation.AcknowledgementsThe project is supported by Wilhelm-Sander-Stiftung (No: 2019.076.1) and by a Roux grant (FKZ: PK37) of the Medical Faculty of the Martin-Luther-University Halle-Wittenberg.ReferencesSubbarayan K, Leisz S, Wickenhauser C, Bethmann D, Massa C, Steven A, Seliger B. Biglycan-mediated upregulation of MHC class I expression in HER-2/neu-transformed cells. Oncoimmunology 2018;7:e1373233.Xu L, Shen SS, Hoshida Y, Subramanian A, Ross K, Brunet JP, Wagner SN, Ramaswamy S, Mesirov JP, Hynes RO. Gene expression changes in an animal melanoma model correlate with aggressiveness of human melanoma metastases. Molecular Cancer Research 2008;6:760-769.

scholarly journals TAP- and tapasin-dependent HLA-E surface expression correlates with the binding of an MHC class I leader peptide

1998 ◽  
Vol 8 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Veronique M. Braud ◽  
David S.J. Allan ◽  
Douglas Wilson ◽  
Andrew J. McMichael
Keyword(s):  
Mhc Class I ◽  
Surface Expression ◽  
Class I ◽  
Leader Peptide

scholarly journals A Distinctive Cytoplasmic Tail Contributes to Low Surface Expression and Intracellular Retention of the Patr-AL MHC Class I Molecule

2015 ◽  
Vol 195 (8) ◽  
pp. 3725-3736 ◽  
Author(s):  
Ana Goyos ◽  
Lisbeth A. Guethlein ◽  
Amir Horowitz ◽  
Hugo G. Hilton ◽  
Michael Gleimer ◽  
...  
Keyword(s):  
Mhc Class I ◽  
Cytoplasmic Tail ◽  
Surface Expression ◽  
Class I ◽  
Mhc Class I Molecule

scholarly journals Modulation of Transporter Associated with Antigen Processing (TAP)-Mediated Peptide Import into the Endoplasmic Reticulum by Flavivirus Infection

2001 ◽  
Vol 75 (12) ◽  
pp. 5663-5671 ◽  
Author(s):  
Frank Momburg ◽  
Arno Müllbacher ◽  
Mario Lobigs
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Surface ◽  
Mhc Class I ◽  
Antigen Processing ◽  
Surface Expression ◽  
Class I ◽  
Peptide Transport ◽  
Cell Surface Expression ◽  
Flavivirus Infection ◽  
Mhc Class I Molecules

ABSTRACT In contrast to many other viruses that escape the cellular immune response by downregulating major histocompatibility complex (MHC) class I molecules, flavivirus infection can upregulate their cell surface expression. Previously we have presented evidence that during flavivirus infection, peptide supply to the endoplasmic reticulum is increased (A. Müllbacher and M. Lobigs, Immunity 3:207–214, 1995). Here we show that during the early phase of infection with different flaviviruses, the transport activity of the peptide transporter associated with antigen processing (TAP) is augmented by up to 50%. TAP expression is unaltered during infection, and viral but not host macromolecular synthesis is required for enhanced peptide transport. This study is the first demonstration of transient enhancement of TAP-dependent peptide import into the lumen of the endoplasmic reticulum as a consequence of a viral infection. We suggest that the increased supply of peptides for assembly with MHC class I molecules in flavivirus-infected cells accounts for the upregulation of MHC class I cell surface expression with the biological consequence of viral evasion of natural killer cell recognition.

scholarly journals Cell Surface Expression of Major Histocompatibility Complex Class I Molecules Is Reduced in Hepatitis C Virus Subgenomic Replicon-Expressing Cells

2003 ◽  
Vol 77 (21) ◽  
pp. 11644-11650 ◽  
Author(s):  
Keith D. Tardif ◽  
Aleem Siddiqui
Keyword(s):  
Major Histocompatibility Complex ◽  
Hepatitis C ◽  
Cell Surface ◽  
Mhc Class I ◽  
Surface Expression ◽  
Protein Glycosylation ◽  
Class I ◽  
Cell Surface Expression ◽  
Histocompatibility Complex ◽  
Mhc Class I Molecules

ABSTRACT The hepatitis C virus (HCV) causes chronic hepatitis in most infected individuals by evading host immune defenses. In this investigation, we show that HCV-infected cells may go undetected in the immune system by suppressing major histocompatibility complex (MHC) class I antigen presentation to cytotoxic T lymphocytes. Cells expressing HCV subgenomic replicons have lower MHC class I cell surface expression. This is due to reduced levels of properly folded MHC class I molecules. HCV replicons induce endoplasmic reticulum (ER) stress (K. Tardif, K. Mori, and A. Siddiqui, J. Virol. 76:7453-7459, 2002), which results from a decline in protein glycosylation. Decreasing protein glycosylation can disrupt protein folding, preventing the assembly of MHC class I molecules. This results in the accumulation of unfolded MHC class I. Therefore, the persistence and pathogenesis of HCV may depend upon the ER stress-mediated interference of MHC class I assembly and cell surface expression.

CLL Cell Proliferation and IL-6 Production Are Regulated by CD160 - MHC Class I Interactions Offering New Therapeutic Targets.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1126-1126
Author(s):  
Jerome Giustiniani ◽  
Catherine Wiseman ◽  
Timothy Farren ◽  
John G. Gribben ◽  
Samir G. Agrawal
Keyword(s):  
Cell Proliferation ◽  
Mhc Class I ◽  
Killer Cell ◽  
Therapeutic Targets ◽  
Cell Types ◽  
Class I ◽  
Cytotoxic Lymphocytes ◽  
Cell Surface Molecule ◽  
Mhc I ◽  
New Therapeutic Targets

Abstract We have previously demonstrated the expression of the Natural Killer cell (NK) marker CD160 in CLL, which is not expressed by normal B cells1. CD160 is a cell surface molecule expressed by human and mouse circulating cytotoxic lymphocytes and exhibits a broad specificity for major histocompatibility complex (MHC) class Ia and Ib molecules 2,3,4. Triggering of CD160 on NK cells leads to cell proliferation and IL-6 production5. IL-6 is a pleiotropic cytokine produced by a variety of cell types, including lymphocytes6. IL-6 expression has been associated with the development of lymphomas7, while higher serum IL-6 levels correlated with shorter survival in CLL8. Without stimulation, CLL cells secreted low basal levels of INF-γ (100pg/ml) and even lower levels of IL-2, IL-4, IL-6, IL-10 and TNF-α (< 50pg/ml) (cytokine bead array, BD Bioscience). Incubation with CL1-R2, an anti-CD160 monoclonal antibody, led to an increase in IL-6 alone (up to 1500pg/ml), which was associated with significant cell proliferation (detected by 3H incorporation) - median 100%, range 50 to 400% (n = 13). Different patterns of response were seen, with all cases showing proliferation to CL1-R2 alone and with the positive control pan anti-immunoglobulin Ab (anti-Ig Ab). In some cases, there was marked synergy between CL1-R2 and anti-Ig Ab. We found that the MEC I and MEC II B-cell lines, derived from a patient with prolymphocytic progression of CLL (DSMZ Institut), express CD160 and show CL1-R2 enhanced proliferation (30 to 55% increase). However, MEC I and MEC II only unregulated IL-10 production (13 to 33% increase). Interestingly, proliferation of CD160Neg Sanchez EBV-immortalized B-cells was inhibited when these cells were incubated with CHO-CD160 transfectants. This inhibition was reversed with an anti-CD160 Ab or anti-MHC-class I W6/32 Ab. The CD160-MHC-I interaction plays a role in CLL biology. MAb engagement of CD160 leads to: a direct proliferative signal; CD160-triggered production of IL-6 and IL-6 mediated effects; inhibition of a constitutive negative signal mediated via MHC-I molecules by their ligand CD160 (supported by the inhibition of the Sanchez cells by CD160 interaction with their MHC class I). The CD160-MHC class I axis represents a new pathway in CLL biology offering new therapeutic targets. This work also suggests a role for targeting the IL-6/IL-6R system in CLL - for example, with humanized anti-IL6 R (Actemra), already used for Castleman’s disease and Rheumatoid Arthritis. Fig 1: cells were incubated 72h in medium only or completed with IgG (control), anti-CD 160 antibody (CL1-R2), anti-IgM, G, A antibody (PAN antibody) or mix with PAN antibody and CL1-R2. For cytokine expriments, supernatants were taken after 24 hours. (left scale: CPM value, right scale: IL-6 concentration in pg/ml) Fig 1:. cells were incubated 72h in medium only or completed with IgG (control), anti-CD 160 antibody (CL1-R2), anti-IgM, G, A antibody (PAN antibody) or mix with PAN antibody and CL1-R2. For cytokine expriments, supernatants were taken after 24 hours. (left scale: CPM value, right scale: IL-6 concentration in pg/ml)

scholarly journals Orf virus interferes with MHC class I surface expression by targeting vesicular transport and Golgi

2012 ◽  
Vol 8 (1) ◽  
pp. 114 ◽  
Author(s):  
Jörg Rohde ◽  
Frederic Emschermann ◽  
Michael R Knittler ◽  
Hanns-Joachim Rziha
Keyword(s):  
Mhc Class I ◽  
Vesicular Transport ◽  
Surface Expression ◽  
Orf Virus ◽  
Class I

MHC class I surface expression in embryo-derived cell lines inducible with peptide or interferon

Nature ◽  
1991 ◽  
Vol 354 (6350) ◽  
pp. 235-238 ◽  
Author(s):  
Elizabeth K. Bikoff ◽  
Leah Jaffe ◽  
Randall K. Ribaudo ◽  
Gillis R. Otten ◽  
Ronald N. Germain ◽  
...  
Keyword(s):  
Mhc Class I ◽  
Cell Lines ◽  
Surface Expression ◽  
Class I
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