جۆری توێژینه‌وه‌: Original Article

نوسه‌ر

Department of Biology, College of Education, University of Garmian, Kalar, Al- Sulaimaniyah, Kurdistan Region, Iraq

پوخته‌

Macrophages as antigen presenting cell (APC) play a vital role in orchestrating immune responses against foreign materials. The activation status of macrophages could be determined by tracking the expression of various cell markers that can be a signal for their immune activity behaviour following cellular stimulation either towards healing or inflammation.  Previously numerous immunofluorescent cell markers have been used for distinguishing between pro-inflammatory macrophage phenotype (M1) and anti-inflammatory macrophage phenotype (M2) qualitatively, although most of those fluorescent cell markers express in both phenotypes. We have developed a new strategy to identify M1 and M2 phenotype quantitatively by using immunofluorescent cell markers. This approach enables the identification of different macrophage functional phenotypes quantitatively, and their degree of polarisation. Macrophages were polarised to M1 and M2 phenotypes by GM-CSF+IFN-γ and M-CSF+IL-4, respectively. Control cells were un-polarised (naïve) macrophages or monocytes were considered as macrophage progeny. For assessing cell polarisation all cell types were stained for nucleus. Also, their surface markers were stained with calprotectin for M1 cells and mannose receptor (MR) for M2 cells, followed by fluorescent microscopy examination. Cell images were analysed using CellProfiler software in order to measure the fluorescent signal intensity of the cell markers, and create a specific profile for each cell type. These profiles formed the basis for M1 and M2 phenotype identification. By using such fluorescent signal parameters we were able to identify M1 and M2 phenotypes effectively and distinguish them from naïve macrophages and monocytes.

وشه‌ بنچینه‌ییه‌كان

Agrawal, H. (2012). Open Journal of Regenerative Medicine 01, 51-59.

Anderson, D. G., Levenberg, S. & Langer, R. (2004). Nature Biotechnology 22, 863-866.

Baitsch, D., Bock, H. H., Engel, T., Telgmann, R., Muller-Tidow, C., Varga, G., Bot, M., Herz, J., Robenek, H., von Eckardstein, A. & Nofer, J. R. (2011). Arteriosclerosis, thrombosis, and vascular biology 31, 1160-1168.

Bartneck, M., Schulte, V. A., Paul, N. E., Diez, M., Lensen, M. C. & Zwadlo-Klarwasser, G. (2010). Acta biomaterialia 6, 3864-3872.

Bradding, P., Feather, I. H., Howarth, P. H., Mueller, R., Roberts, J. A., Britten, K., Bews, J. P., Hunt, T. C., Okayama, Y., Heusser, C. H. & et al. (1992). The Journal of experimental medicine 176, 1381-1386.

Chinetti-Gbaguidi, G., Baron, M., Bouhlel, M. A., Vanhoutte, J., Copin, C., Sebti, Y., Derudas, B., Mayi, T., Bories, G., Tailleux, A., Haulon, S., Zawadzki, C., Jude, B. & Staels, B. (2011). Circ Res 108, 985-995.

Davis, M. J., Tsang, T. M., Qiu, Y., Dayrit, J. K., Freij, J. B., Huffnagle, G. B. & Olszewski, M. A. (2013). mBio 4, e00264-00213.

De Paoli, F., Staels, B. & Chinetti-Gbaguidi, G. (2014). Circulation Journal advpub.

Edin, S., Wikberg, M. L., Dahlin, A. M., Rutegard, J., Oberg, A., Oldenborg, P. A. & Palmqvist, R. (2012). Plos One 7, e47045.

El Kasmi, K. C., Qualls, J. E., Pesce, J. T., Smith, A. M., Thompson, R. W., Henao-Tamayo, M., Basaraba, R. J., Konig, T., Schleicher, U., Koo, M. S., Kaplan, G., Fitzgerald, K. A., Tuomanen, E. I., Orme, I. M., Kanneganti, T. D., Bogdan, C., Wynn, T. A. & Murray, P. J. (2008). Nature immunology 9, 1399-1406.

Fleming, B. D. & Mosser, D. M. (2011). Eur J Immunol 41, 2498-2502.

Garcia, S., Krausz, S., Ambarus, C. A., Fernandez, B. M., Hartkamp, L. M., van Es, I. E., Hamann, J., Baeten, D. L., Tak, P. P. & Reedquist, K. A. (2014). Plos One 9.

Goerdt, S. & Orfanos, C. E. (1999). Immunity 10, 137-142.

Hagert, C., Sareila, O., Kelkka, T., Jalkanen, S. & Holmdahl, R. (2018). Frontiers in immunology 9, 114.

Hamilton, J. A. (2002). Trends in immunology 23, 403-408.

Hamilton, J. A. (2008). Nat Rev Immunol 8, 533-544.

Hao, N. B., Lu, M. H., Fan, Y. H., Cao, Y. L., Zhang, Z. R. & Yang, S. M. (2012). Clinical & developmental immunology 2012, 948098.

Hofkens, W., Storm, G., van den Berg, W. & van Lent, P. (2011). Ann Rheum Dis 70, A40-A40.

Hook, A. L., Chang, C. Y., Yang, J., Scurr, D. J., Langer, R., Anderson, D. G., Atkinson, S., Williams, P., Davies, M. C. & Alexander, M. R. (2012). Jove-J Vis Exp.

Hsu, K., Champaiboon, C., Guenther, B. D., Sorenson, B. S., Khammanivong, A., Ross, K. F., Geczy, C. L. & Herzberg, M. C. (2009). Anti-inflammatory & anti-allergy agents in medicinal chemistry 8, 290-305.

Huang, R., Wang, X., Zhou, Y. & Xiao, Y. (2017). Bone research 5, 17019.

Kratky, W., Sousa, C. R. E., Oxenius, A. & Sporria, R. (2011). Proc Natl Acad Sci U S A 108, 17414-17419.

Lee, H. S., Stachelek, S. J., Tomczyk, N., Finley, M. J., Composto, R. J. & Eckmann, D. M. (2013). J Biomed Mater Res A 101, 203-212.

Leitinger, N. & Schulman, I. G. (2013). Arterioscl Throm Vas 33, 1120-1126.

Mantovani, A. (2006). Blood 108, 408-409.

Mantovani, A., Sica, A., Sozzani, S., Allavena, P., Vecchi, A. & Locati, M. (2004). Trends in immunology 25, 677-686.

Martinez, F. O. & Gordon, S. (2014). F1000prime reports 6, 13.

McWhorter, F. Y., Wang, T. T., Nguyen, P., Chung, T. & Liu, W. F. (2013). P Natl Acad Sci USA 110, 17253-17258.

Mills, C. D., Kincaid, K., Alt, J. M., Heilman, M. J. & Hill, A. M. (2000). J Immunol 164, 6166-6173.

Mosser, D. M. & Edwards, J. P. (2008). Nat Rev Immunol 8, 958-969.

Murray, P. J., Allen, J. E., Biswas, S. K., Fisher, E. A., Gilroy, D. W., Goerdt, S., Gordon, S., Hamilton, J. A., Ivashkiv, L. B., Lawrence, T., Locati, M., Mantovani, A., Martinez, F. O., Mege, J. L., Mosser, D. M., Natoli, G., Saeij, J. P., Schultze, J. L., Shirey, K. A., Sica, A., Suttles, J., Udalova, I., van Ginderachter, J. A., Vogel, S. N. & Wynn, T. A. (2014). Immunity 41, 14-20.

Murray, P. J. & Wynn, T. A. (2011). Nat Rev Immunol 11, 723-737.

Pelegrin, P. & Surprenant, A. (2009). The EMBO journal 28, 2114-2127.

Porcheray, F., Viaud, S., Rimaniol, A. C., Leone, C., Samah, B., Dereuddre-Bosquet, N., Dormont, D. & Gras, G. (2005). Clinical and experimental immunology 142, 481-489.

Rostam, H. M., Reynolds, P. M., Alexander, M. R., Gadegaard, N. & Ghaemmaghami, A. M. (2017). Scientific reports 7, 3521.

Rostam, H. M., Singh, S., Salazar, F., Magennis, P., Hook, A., Singh, T., Vrana, N. E., Alexander, M. R. & Ghaemmaghami, A. M. (2016). Immunobiology 221, 1237-1246.

Rostam, H. M., Singh, S., Vrana, N. E., Alexander, M. R. & Ghaemmaghami, A. M. (2015). Biomaterials science 3, 424-441.

Sindrilaru, A., Peters, T., Wieschalka, S., Baican, C., Baican, A., Peter, H., Hainzl, A., Schatz, S., Qi, Y., Schlecht, A., Weiss, J. M., Wlaschek, M., Sunderkotter, C. & Scharffetter-Kochanek, K. (2011). The Journal of clinical investigation 121, 985-997.

Singh, S., Awuah, D., Rostam, H. M., Emes, R. D., Kandola, N. K., Onion, D., Htwe, S. S., Rajchagool, B., Cha, B. H., Kim, D., Tighe, P. J., Vrana, N. E., Khademhosseini, A. & Ghaemmaghami, A. (2017). Acs Biomater Sci Eng 3, 969-978.

Sutterwala, F. S., Noel, G. J., Clynes, R. & Mosser, D. M. (1997). J Exp Med 185, 1977-1985.

Taylor, M., Urquhart, A. J., Zelzer, M., Davies, M. C. & Alexander, M. R. (2007). Langmuir 23, 6875-6878.

Urquhart, A. J., Anderson, D. G., Taylor, M., Alexander, M. R., Langer, R. & Davies, M. C. (2007). Adv Mater 19, 2486-+.

Vereyken, E. J., Heijnen, P. D., Baron, W., de Vries, E. H., Dijkstra, C. D. & Teunissen, C. E. (2011). Journal of neuroinflammation 8, 58.

Verreck, F. A. W., de Boer, T., Langenberg, D. M. L., Hoeve, M. A., Kramer, M., Vaisberg, E., Kastelein, R., Kolk, A., de Waal-Malefyt, R. & Ottenhoff, T. H. M. (2004). P Natl Acad Sci USA 101, 4560-4565.

Willenborg, S., Lucas, T., van Loo, G., Knipper, J. A., Krieg, T., Haase, I., Brachvogel, B., Hammerschmidt, M., Nagy, A., Ferrara, N., Pasparakis, M. & Eming, S. A. (2012). Blood 120, 613-625.

Xia, C., Braunstein, Z., Toomey, A. C., Zhong, J. & Rao, X. (2018). Frontiers in immunology 8.

Xue, J., Schmidt, S. V., Sander, J., Draffehn, A., Krebs, W., Quester, I., De Nardo, D., Gohel, T. D., Emde, M., Schmidleithner, L., Ganesan, H., Nino-Castro, A., Mallmann, M. R., Labzin, L., Theis, H., Kraut, M., Beyer, M., Latz, E., Freeman, T. C., Ulas, T. & Schultze, J. L. (2014). Immunity 40, 274-288.