Comparison of Interferon-gamma Secretion by Stimulated NK Cells and T cells from Healthy Subjects

Gyu-Dae An; Kyeong-Hee Kim; Hyeon-Ho Lim; Min-Chan Kim; Sang Yeob Lee

doi:10.3343/lmo.2018.8.1.15

Journal List > Lab Med Online > v.8(1) > 1057377

Go to TopGo to Top Go to BottomGo to Bottom

TOOLS

An, Kim, Lim, Kim, and Lee: Comparison of Interferon-gamma Secretion by Stimulated NK Cells and T cells from Healthy Subjects

Original Article

Lab Med Online 2018;8(1):15-18.

Published online: 19 January 2018

DOI: https://doi.org/10.3343/lmo.2018.8.1.15

Comparison of Interferon-gamma Secretion by Stimulated NK Cells and T cells from Healthy Subjects

Gyu-Dae An, M.D.¹, Kyeong-Hee Kim, M.D.¹, Hyeon-Ho Lim, M.D.¹, Min-Chan Kim, M.D.², Sang Yeob Lee, M.D.³

¹Departments of Laboratory Medicine, Busan, Korea

²Departments of Surgery, Busan, Korea

³ Departments of Rheumatology, Dong-A University College of Medicine, Busan, Korea

Corresponding author: Kyeong-Hee Kim Department of Laboratory Medicine, Dong-A University College of Medicine, Dong-A University Hospital, 26 Daeshingongwon-ro, Seo-gu, Busan 49202, Korea Tel: +82-51-240-2850, Fax: +82-51-255-9366, E-mail: progreen@dau.ac.kr

Received 20 February 2017 Revised 7 June 2017 Accepted 21 June 2017

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Interferon-γ (IFN-γ) is an important cytokine produced by natural killer (NK) cells and T cells in response to various stimuli. The levels of IFN-γ secreted after stimulation of NK cells using a recombinant cytokine is represented as one of functions of NK cells. Recently, a method for evaluating NK cell activity in whole blood samples was developed. The levels of IFN-γ secreted after NK cell stimulation with PROMOCA^TM (ATGen, Korea) and T cell stimulation with phytohemagglutinin (PHA) were compared using two different commercial kits: NK Vue Gold (ATGen, Korea) and QuantiFER-ON-TB Gold In-Tube (Cellestis, Australia). Participants included 43 healthy individuals. Whole blood samples were incubated with either PROMOCA, a recombinant cytokine that specifically activates NK cells, or with PHA. IFN-γ levels in the supernatants were measured by ELISA. The level of IFN-γ by PROMOCA stimulation (PROMOCA IFN-γ) was more varied than that by stimulation with PHA (PHA IFN-γ) (median 1,544.4 pg/mL [range 193.7–2,530.9] vs. median 2,470.1 pg/mL [2,250.1–2,874.4] P=0.0001). The median of PHA IFN-γ/PROMOCA IFN-γ ratio was 1.9 (1.1–12.4). There was a significant difference in levels of IFN-γ secreted after stimulation with PROMOCA or PHA in the healthy population.

Keywords: Natural killer cells, Interferon-gamma, Phytohemagglutinin, ELISA, PROMOCA

REFERENCES

1.Vivier E., Raulet DH., Moretta A., Caligiuri MA., Zitvogel L., Lanier LL, et al. Innate or adaptive immunity? The example of natural killer cells. Science. 2011. 331:44–9.

2.Brunner KT., Mauel J., Cerottini JC., Chapuis B. Quantitative assay of the lytic action of immune lymphoid cells on 51-Cr-labelled allogeneic target cells in vitro; inhibition by isoantibody and by drugs. Immunology. 1968. 4:181–96.

3.Jang YY., Cho D., Kim SK., Shin DJ., Park MH., Lee JJ, et al. An improved fow cytometry-based natural killer cytotoxicity assay involving cal-cein AM staining of effector cells. Ann Clin Lab Sci. 2012. 42:42–9.

4.Yamashita M., Kitano S., Aikawa H., Kuchiba A., Hayashi M., Yamamoto N, et al. A novel method for evaluating antibody-dependent cell-mediated cytotoxicity by fowcytometry using cryopreserved human peripheral blood mononuclear cells. Sci Rep. 2016. 6:19772.

5.Alter G., Malenfant JM., Altfeld M. CD107a as a functional marker for the identifcation of natural killer cell activity. J Immunol Methods. 2004. 294:15–22.

6.Lee SB., Cha J., Kim IK., Yoon JC., Lee HJ., Park SW, et al. A high-throughput assay of NK cell activity in whole blood and its clinical application. Biochem Biophys Res Commun. 2014. 445:584–90.

7.Nowell PC. Phytohemagglutinin: an initiator of mitosis in cultures of normal human leukocytes. Cancer Res. 1960. 20:462–6.

8.Wheelock EF. Interferon-like virus inhibitor induced in human leukocytes by phytohemagglutinin. Science. 1965. 149:310–1.

9.Chilson OP., Boylston AW., Crumpton MJ. Phaseolus vulgaris phytohaemagglutinin (PHA) binds to the human T lymphocyte antigen receptor. EMBO J. 1984. 3:3239–45.

10.Billiau A., Matthys P. Interferon-gamma: a historical perspective. Cytokine Growth Factor Rev. 2009. 20:97–113.

11.Woo KS., Choi JL., Kim BR., Kim JE., Kim BG., Lee H, et al. Signifcance of interferon-gamma response to mitogen in serial QuantiFERON-TB Gold In-Tube assay of routine laboratory practice. Clin Chim Acta. 2014. 430:79–83.

12.Lange B., Vavra M., Kern WV., Wagner D. Indeterminate results of a tuberculosis-specifc interferon gamma release assay in immunocompromised patients. Eur Respir J. 2010. 35:1179–82.

13.Fabre V., Shoham S., Page KR., Shah M. High proportion of indeterminate QuantiFERON-TB Gold In-Tube results in an inpatient population is related to host factors and preanalytical steps. Open Forum In-fect Dis. 2014. 1:ofu088.

14.Aichelburg MC., Tittes J., Breitenecker F., Reiberger T., Kohrgruber N., Rieger A. Prognostic value of indeterminate IFN-γ release assay results in HIV-1 infection. J Clin Microbiol. 2012. 50:2767–9.

15.Banaei N., Gaur RL., Pai M. Interferon gamma release assays for latent tuberculosis: What are the sources of variability? J Clin Microbiol. 2016. 54:845–50.

16.De Maria A., Bozzano F., Cantoni C., Moretta L. Revisiting human natural killer cell subset function revealed cytolytic CD56(dim)CD16+ NK cells as rapid producers of abundant IFN-gamma on activation. Proc Natl Acad Sci USA. 2011. 108:728–32.

17.Imai K., Matsuyama S., Miyake S., Suga K., Nakachi K. Natural cytotoxic activity of peripheral-blood lymphocytes and cancer incidence: an 11-year follow-up study of a general population. Lancet. 2000. 356:1795–9. 18. Lim YA, Kim SS, Cho SW, Cheong JY. Evaluation of the effectiveness of NK Vue Gold kit in patients with chronic hepatitis B. J Lab Med Qual Assur 2016;38: 151-8.

18.Lim YA., Kim SS., Cho SW., Cheong JY. Evaluation of the effectiveness of NK Vue Gold kit in patients with chronic hepatitis B. J Lab Med Qual Assur. 2016. 38:151–8.

19.Hou H., Mao L., Wang J., Liu W., Lu Y., Yu J, et al. Establishing the reference intervals of NK cell functions in healthy adults. Hum Immunol. 2016. 77:637–42.

20.Koo KC., Shim DH., Yang CM., Lee SB., Kim SM., Shin TY, et al. Reduction of the CD16(-)CD56bright NK cell subset precedes NK cell dysfunction in prostate cancer. PLoS One. 2013. 8:e78049.

21.Dahlberg CI., Sarhan D., Chrobok M., Duru AD., Alici E. Natural killer cell-based therapies targeting cancer: possible strategies to gain and sustain anti-tumor activity. Front Immunol. 2015. 6:605.

22.Guillerey C., Huntington ND., Smyth MJ. Targeting natural killer cells in cancer immunotherapy. Nat Immunol. 2016. 17:1025–36.

Fig. 1.

Comparison of levels of IFN-γ secretion between PROMOCA stimulation and PHA stimulation (P =0.0001 by Mann-Whitney test). Horizontal bar indicates 95% confidence interval for the median.

Fig. 2.

Pearson correlation for levels of IFN-γ secreted after PROMOCA stimulation and PHA stimulation.

TOOLS

Similar articles