Download PDF
Drug Res (Stuttg) 2014; 64(6): 281-286
DOI: 10.1055/s-0033-1358465
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Augmenting the Expression of NKp44 Molecule and the Natural Killer Activity in Peripheral Blood Mononuclear Cells from Patients with Malignant Colorectal Carcinoma

A. Sheikhi
1   Department of Immunology, Faculty of Medicine, Dezful University of Medical Sciences, Dezful, Iran
2   Department of Immunology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
,
K. Saadati
3   Department of Surgery, Zanjan University of Medical Sciences, Zanjan, Iran
,
A. Jafarzadeh
4   Department of Immunology, Faculty of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
5   Department of Immunology, Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran
,
H. Karimi
6   Department of Internal Medicine, Dezful University of Medical Sciences, Dezful, Iran
,
N. Mousavinasab
7   Department of Social Medicine, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
› Author Affiliations
Further Information

Publication History

received 18 July 2013

accepted 21 September 2013

Publication Date:
23 October 2013 (online)

Also available at
    Permissions and Reprints

Abstract

Objective: NKp44 and NKG2D are of the main NK activating receptors involved in recognition and killing of tumors. Here we studied the stimulatory effects of PHA and/or K562 cell line on induction of NKp44 and NKG2D expression and the NK activity of PBMCs from patients with colorectal carcinoma (CRC).

Materials and Methods: Peripheral blood samples were collected from 10 patients with CRC. The peripheral blood mononuclear cells (PBMCs) from each patient received a single stimulation with PHA or double stimulation with PHA and irradiated K562 cell line (iK562). The expression of CD56, NKG2D and NKp44 were detected by flowcytometry. The NK activity of PBMCs against a colorectal carcinoma cell line named as SW742 was determined with 51Cr-release assay.

Results: Double stimulation of PBMCs with PHA+iK562 significantly augmented the number CD56+ cells compared to PHA alone and non-stimulated PBMCs (P<0.000, P<0.0000; respectively). A single stimulation of PBMCs with PHA resulted in an enhancement in NKG2D and NKp44 expression from 16.6±3.3% (for non-stimulated PBMCs) to 42±5.6% and 48.1±3.8% respectively (p<0.05). Double stimulation of PBMCs augmented the NKp44 expression significantly in comparison with single stimulation with PHA (73.6±12%, p<0.05). Double stimulation of PBMCs significantly enhanced the NK activity against SW742 target cells compared to single stimulation with PHA (p<0.05).

Discussion and Conclusion: Our results demonstrated that the mitogen and iK562 exposure to PBMCs can significantly improve NK activity which is co-related to the higher expression of NKp44 and NKG2D. These data may help to improve cancer immunotherapy protocols.

Key words

natural killer activity - immunomodulation - K562 - NKp44 - NKG2D - CRC
 

  • References

  • 1 Huang B, Sikorski R, Sampath P et al. Modulation of NKG2D-ligand cell surface expression enhances immune cell therapy of cancer. J Immunother 2011; 34: 289-296
    CrossrefPubMedSearch in Google Scholar
  • 2 Linn YC, Hui KM. Cytokine-induced killer cells: NK-like T cells with cytotolytic specificity against leukemia. Leuk Lymphoma 2003; 44: 1457-1462
    CrossrefPubMedSearch in Google Scholar
  • 3 Bacon CM, Lawry J, Rees RC. TNF alpha protects SW742 human colon carcinoma cells against non-MHC-restricted cytolysis. Immunol Lett 1993; 38: 215-222
    CrossrefPubMedSearch in Google Scholar
  • 4 Tallerico R, Todaro M, Di Franco S et al. Human NK cells selective targeting of colon cancer-initiating cells: a role for natural cytotoxicity receptors and MHC class I molecules. J Immunol 2013; 190: 2381-2390
    CrossrefPubMedSearch in Google Scholar
  • 5 Moretta L, Biassoni R, Bottino C et al. Natural killer cells: a mystery no more. Scand J Immunol 2002; 55: 229-232
    CrossrefPubMedSearch in Google Scholar
  • 6 Pende D, Parolini S, Pessino A et al. Identification and molecular characterization of NKp30, a novel triggering receptor involved in natural cytotoxicity mediated by human natural killer cells. J Exp Med 1999; 190: 1505-1516
    CrossrefPubMedSearch in Google Scholar
  • 7 Diefenbach A, Jamieson AM, Liu SD et al. Ligands for the murine NKG2D receptor: expression by tumor cells and activation of NK cells and macrophages. Nat Immunol 2000; 1: 119-126
    CrossrefPubMedSearch in Google Scholar
  • 8 Mandelboim O, Lieberman N, Lev M et al. Recognition of haemagglutinins on virus-infected cells by NKp46 activates lysis by human NK cells. Nature 2001; 409: 1055-1060
    CrossrefPubMedSearch in Google Scholar
  • 9 Arnon TI, Lev M, Katz G et al. Recognition of viral hemagglutinins by NKp44 but not by NKp30. Eur J Immunol 2001; 31: 2680-2689
    CrossrefPubMedSearch in Google Scholar
  • 10 Arnon TI, Achdout H, Lieberman N et al. The mechanisms controlling the recognition of tumor and virus infected cells by NKp46. Blood 2004; 103: 664-672
    CrossrefPubMedSearch in Google Scholar
  • 11 Arnon TI, Achdout H, Levi O et al. Inhibition of the NKp30 activating receptor by pp65 of human cytomegalovirus. Nat Immunol 2005; 6: 515-523
    CrossrefPubMedSearch in Google Scholar
  • 12 Moretta L, Bottino C, Cantoni C et al. Human natural killer cell function and receptors. Curr Opin Pharmacol 2001; 1: 387-391 Review
    CrossrefPubMedSearch in Google Scholar
  • 13 Moretta L, Moretta A. Killer immunoglobulin-like receptors. Curr Opin Immunol 2004; 16: 626-633 Review
    CrossrefPubMedSearch in Google Scholar
  • 14 Byrd A, Hoffmann SC, Jarahian M et al. Expression analysis of the ligands for the Natural Killer cell receptors NKp30 and NKp44. PLoS One 2007; 2: e1339
    CrossrefPubMedSearch in Google Scholar
  • 15 Pende D, Cantoni C, Rivera P et al. Role of NKG2D in tumor cell lysis mediated by human NK cells: cooperation with natural cytotoxicity receptors and capability of recognizing tumors of nonepithelial origin. Eur J Immunol 2001; 31: 1076-1086
    CrossrefPubMedSearch in Google Scholar
  • 16 Jemal A, Siegel R, Ward E et al. Cancer statistics 2006. CA Cancer J Clin 2006; 56: 106-130
    Search in Google Scholar
  • 17 Iovino F, Meraviglia S, Spina M et al. Immunotherapy targeting colon cancer stem cells. Immunotherapy 2011; 3: 97-106
    CrossrefPubMedSearch in Google Scholar
  • 18 Zhang J, Zhang JK, Zhuo SH et al. Effect of a cancer vaccine prepared by fusions of hepatocarcinoma cells with dendritic cells. World J Gastroenterol 2001; 7: 690-694
    PubMedSearch in Google Scholar
  • 19 Guven H, Gilljam M, Chambers B et al. Expansion of natural killer (NK) and natural killer-like T (NKT)-cell populations derived from patients with B-chronic lymphocytic leukemia (B-CLL): a potential source for cellular immunotherapy. Leukemia 2003; 17: 1973-1980
    CrossrefPubMedSearch in Google Scholar
  • 20 Cho D, Campana D. Expansion and activation of natural killer cells for cancer immunotherapy. Korean J Lab Med 2009; 29: 89-96 Review
    CrossrefPubMedSearch in Google Scholar
  • 21 Sheikhi A, Saadati K, Salmani R et al. In vitro modulation of natural killer activity of human peripheral blood mononuclear cells against prostate tumor cell line. Immunopharmacol Immunotoxicol 2011; 33: 700-708
    CrossrefPubMedSearch in Google Scholar
  • 22 Karre K, Ljunggren HG, Piontek G et al. Selective rejection of H-2-deficient lymphoma variants suggests alternative immune defence strategy. Nature 1986; 319: 675-678
    CrossrefPubMedSearch in Google Scholar
  • 23 Long EO. Regulation of immune responses through inhibitory receptors. Annu Rev Immunol 1999; 17: 875-904
    CrossrefPubMedSearch in Google Scholar
  • 24 Lanier LL. Natural killer cell receptor signaling. Curr Opin Immunol 2003; 15: 308-314
    CrossrefPubMedSearch in Google Scholar
  • 25 Marin R, Ruiz-Cabello F, Pedrinaci S et al. Analysis of HLA-E in human tumours. Immunogenetics 2003; 54: 767-775
    PubMedSearch in Google Scholar
  • 26 Levy EM, Bianchini M, Von Euw EM et al. Human leukocyte antigen E protein is overexpressed in primary human colorectal cancer. Int J Oncol 2008; 32: 633-641
    PubMedSearch in Google Scholar
  • 27 Sheikhi AK, Amirghofran Z. Influence of two recombinant sHLA molecules on NK cell-mediated lysis of cells with different HLA class I expression. Archives of Iranian Medicine 2003; 6: 282-288
    PubMedSearch in Google Scholar
  • 28 Haas GP, Solomon D, Rosenberg SA. Tumor-infiltrating lymphocytes from nonrenal urological malignancies. Cancer Immunol Immunother 1990; 30: 342-350
    CrossrefPubMedSearch in Google Scholar
  • 29 Ruggeri L, Capanni M, Casucci M et al. Role of natural killer cell alloreactivity in HLA-mismatched hematopoietic stem cell transplantation. Blood 1999; 94: 333-339
    PubMedSearch in Google Scholar
  • 30 Ruggeri L, Capanni M, Urbani E et al. Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants. Science 2002; 295: 2097-2100
    CrossrefPubMedSearch in Google Scholar
  • 31 Karre K. A perfect mismatch. Science 2002; 295: 2029-2031
    CrossrefPubMedSearch in Google Scholar
  • 32 Grönberg A, Ferm MT, Ng J et al. IFN-gamma treatment of K562 cells inhibits natural killer cell triggering and decreases the susceptibility to lysis by cytoplasmic granules from large granular lymphocytes. J Immunol 1988; 140: 4397-4402
    PubMedSearch in Google Scholar
  • 33 Stark GR, Friedman RL, McMahon M et al. Induction of human mRNAs by interferon. Philos Trans R Soc Lond B Biol Sci 1984; 307: 227-230
    CrossrefPubMedSearch in Google Scholar