Микровезикулы естественных киллеров линии NK-92 влияют на фенотип и функции эндотелиальных клеток линии EA.Hy926

Микровезикулы (МВ) — субклеточные структуры размером от 100 до 1000 нм, продуцируемые клетками в состоянии покоя и активации. МВ могут передавать молекулы клеткам-мишеням, регулировать физиологические процессы, участвовать в патологиях. Микровезикулы лейкоцитарного происхождения, в частности МВ NK-клеток, остаются наименее изученной популяцией МВ. NK-клетки способны изменять функциональную активность эндотелиальных клеток (ЭК), участвуют в регуляции ангиогенеза. Недостаточно изучена способность МВ NK-клеток влиять на функциональное состояние ЭК. Целью настоящего исследования явилось изучение влияния МВ, образуемых естественными киллерами линии NK-92, на фенотип, активность каспаз, пролиферацию и миграцию ЭК линии EA.Hy926. ЭК культивировали в присутствии МВ клеток линии NK-92. При помощи проточной цитофлуориметрии оценивали изменение фенотипа ЭК, передачу внутриклеточного белка из МВ в ЭК, относительную гибель ЭК. При помощи Western blot analysis оценивали экспрессию гранзима B в NK-клетках и их МВ, появление гранзима B в ЭК, экспрессию каспаз, Erk, AKT в ЭК. Также оценивали пролиферацию и миграцию ЭК в присутствии МВ клеток линии NK-92. Установлено значимое различие протеомных профилей клеток линии NK-92 и образуемых ими МВ. Контакт ЭК с МВ клеток линии NK-92 сопровождается развитием следующих событий: 1) экспрессией в ЭК гранзима В; 2) активацией каспазы-9, каспазы-3 и частичной гибелью ЭК; 3) появлением на ЭК панлейкоцитарного маркера CD45; 4) снижением экспрессии CD105 и повышением экспрессии CD34 и CD54; 5) ингибированием миграции ЭК. Передача эндотелиальным клеткам Erk, но не AKT, в составе МВ клеток линии NK-92 в концентрации в 10 раз ниже концентрации, вызывающей гибель ЭК, способствует повышению пролиферации ЭК.

1. Andreu Z., Yanez-Mo M. Tetraspanins in extracellular vesicle formation and function. Front. Immunol., 2014, Vol. 5, 442. doi: 10.3389/fimmu.2014.00442.

2. Ardoin S.P., Shanahan J.C., Pisetsky D.S. The role of microparticles in inflammation and thrombosis. Scand. J. Immunol., 2007, Vol. 66, no. 2-3, pp. 159-165.

3. Ashkenazi A., Salvesen G. Regulated cell death: signaling and mechanisms. Annu. Rev. Cell Dev. Biol., 2014, Vol. 30, pp. 337-356.

4. Ashton S.V., Whitley G.S., Dash P.R., Wareing M., Crocker I.P., Baker P.N., Cartwright J.E. Uterine spiral artery remodeling involves endothelial apoptosis induced by extravillous trophoblasts through Fas/FasL interactions. Arterioscler. Thromb. Vasc. Biol., 2005, Vol. 25, no. 1, pp. 102-108.

5. Aubrey B.J., Kelly G.L., Janic A., Herold M.J., Strasser A. How does p53 induce apoptosis and how does this relate to p53-mediated tumour suppression? Cell Death Differ., 2018, Vol. 25, no. 1, pp. 104-113.

6. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 1976, Vol. 72, pp. 248-254.

7. Budaj M., Poljak Z., Duris I., Kasko M., Imrich R., Kopani M., Maruscakova L., Hulin I. Microparticles: a component of various diseases. Pol. Arch. Med. Wewn., 2012, Vol. 122, Suppl. 1, pp. 24-29.

8. Burger D., Schock S., Thompson C.S., Montezano A.C., Hakim A.M., Touyz R.M. Microparticles: biomarkers and beyond. Clin. Sci. (Lond.), 2013, Vol. 124, no. 7, pp. 423-441.

9. Cantarella G., di Benedetto G., Ribatti D., Saccani-Jotti G., Bernardini R. Involvement of caspase 8 and c-FLIPL in the proangiogenic effects of the tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). FEBS J., 2014, Vol. 281, no. 5, pp. 1505-1513.

10. Chazara O., Xiong S., Moffett A. Maternal KIR and fetal HLA-C: a fine balance. J. Leukoc. Biol., 2011, Vol. 90, no. 4, pp. 703-716.

11. Colombo M., Raposo G., Thery C. Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu. Rev. Cell Dev. Biol., 2014, Vol. 30, pp. 255-289.

12. Cooper M.A., Fehniger T.A., Caligiuri M.A. The biology of human natural killer-cell subsets. Trends Immunol., 2001, Vol. 22, no. 11, pp. 633-640.

13. Dasgupta S.K., Abdel-Monem H., Niravath P., Le A., Bellera R.V, Langlois K., Nagata S., Rumbaut R.E., Thiagarajan P. Lactadherin and clearance of platelet-derived microvesicles. Blood, 2009, Vol. 113, no. 6, pp. 1332-1339.

14. Delia D., Lampugnani M.G., Resnati M., Dejana E., Aiello A., Fontanella E., Soligo D., Pierotti M.A., Greaves M.F. CD34 expression is regulated reciprocally with adhesion molecules in vascular endothelial cells in vitro. Blood, 1993, Vol. 81, no. 4, pp. 1001-1008.

15. Distler J.H., Huber L.C., Gay S., Distler O., Pisetsky D.S. Microparticles as mediators of cellular cross-talk in inflammatory disease. Autoimmunity, 2006, Vol. 39, no. 8, pp. 683-690.

16. Dondero A., Casu B., Bellora F., Vacca A., de Luisi A., Frassanito M.A., Cantoni C., Gaggero S., Olive D., Moretta A., Bottino C., Castriconi R. NK cells and multiple myeloma-associated endothelial cells: molecular interactions and influence of IL-27. Oncotarget, 2017, Vol. 8, no. 21, pp. 35088-35102.

17. Dragovic R.A., Collett G.P., Hole P., Ferguson D.J., Redman C.W., Sargent I.L., Tannetta D.S. Isolation of syncytiotrophoblast microvesicles and exosomes and their characterisation by multicolour flow cytometry and fluorescence Nanoparticle Tracking Analysis. Methods, 2015, Vol. 87, pp. 64-74.

18. Edgell C.J., McDonald C.C., Graham J.B. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc. Natl. Acad. Sci. USA, 1983, Vol. 80, no. 12, pp. 3734-3737.

19. el Costa H., Tabiasco J., Berrebi A., Parant O., Aguerre-Girr M., Piccinni M.P., le Bouteiller P. Effector functions of human decidual NK cells in healthy early pregnancy are dependent on the specific engagement of natural cytotoxicity receptors. J. Reprod. Immunol., 2009, Vol. 82, no. 2, pp. 142-147.

20. Evans-Osses I., Reichembach L.H., Ramirez M.I. Exosomes or microvesicles? Two kinds of extracellular vesicles with different routes to modify protozoan-host cell interaction. Parasitol. Res., 2015, Vol. 114, no. 10, pp. 3567-3575.

21. Fraser R., Whitley G.S., Thilaganathan B., Cartwright J.E. Decidual natural killer cells regulate vessel stability: implications for impaired spiral artery remodelling. J. Reprod. Immunol., 2015, Vol. 110, pp. 54-60.

22. Gojova A., Barakat A.I. Vascular endothelial wound closure under shear stress: role of membrane fluidity and flow-sensitive ion channels. J. Appl. Physiol. (1985), 2005, Vol. 98, no. 6, pp. 2355-2362.

23. Gong J.H., Maki G., Klingemann H.G. Characterization of a human cell line (NK-92) with phenotypical and functional characteristics of activated natural killer cells. Leukemia, 1994, Vol. 8, no. 4, pp. 652-658.

24. Halim A.T., Ariffin N.A., Azlan M. Review: the Multiple roles of monocytic microparticles. Inflammation, 2016, Vol. 39, no. 4, pp. 1277-1284.

25. Hanna J., Goldman-Wohl D., Hamani Y., Avraham I., Greenfield C., Natanson-Yaron S., Prus D., Cohen-Daniel L., Arnon T.I., Manaster I., Gazit R., Yutkin V, Benharroch D., Porgador A., Keshet E., Yagel S., Mandelboim O. Decidual NK cells regulate key developmental processes at the human fetal-maternal interface. Nat. Med., 2006, Vol. 12, no. 9, pp. 1065-1074.

26. Hanna J., Wald O., Goldman-Wohl D., Prus D., Markel G., Gazit R., Katz G., Haimov-Kochman R., Fujii N., Yagel S., Peled A., Mandelboim O. CXCL12 expression by invasive trophoblasts induces the specific migration of CD16- human natural killer cells. Blood, 2003, Vol. 102, no. 5, pp. 1569-1577.

27. Hemler M.E. Tetraspanin proteins mediate cellular penetration, invasion, and fusion events and define a novel type of membrane microdomain. Annu. Rev. Cell Dev. Biol., 2003, Vol. 19, pp. 397-422.

28. Imam J.S., Buddavarapu K., Lee-Chang J.S., Ganapathy S., Camosy C., Chen Y., Rao M.K. MicroRNA-185 suppresses tumor growth and progression by targeting the Six1 oncogene in human cancers. Oncogene, 2010, Vol. 29, no. 35, pp. 4971-4979.

29. Kalkunte S.S., Mselle T.F., Norris W.E., Wira C.R., Sentman C.L., Sharma S. Vascular endothelial growth factor C facilitates immune tolerance and endovascular activity of human uterine NK cells at the maternal-fetal interface. J. Immunol., 2009, Vol. 182, no. 7, pp. 4085-4092.

30. Kalra H., Drummen G.P., Mathivanan S. Focus on extracellular vesicles: introducing the next small big thing. Int. J. Mol. Sci, 2016, Vol. 17, no. 2, 170. doi: 10.3390/ijms17020170.

31. Kawakami A., Hida A., Yamasaki S., Miyashita T., Nakashima K., Tanaka F., Ida H., Furuyama M., Migita K., Origuchi T., Eguchi K. Modulation of the expression of membrane-bound CD54 (mCD54) and soluble form of CD54 (sCD54) in endothelial cells by glucosyl transferase inhibitor: possible role of ceramide for the shedding of mCD54. Biochem. Biophys. Res. Commun., 2002, Vol. 296, no. 1, pp. 26-31.

32. Kawauchi K., Ihjima K., Yamada O. IL-2 increases human telomerase reverse transcriptase activity transcriptionally and posttranslationally through phosphatidylinositol 3’-kinase/Akt, heat shock protein 90, and mammalian target of rapamycin in transformed NK cells. J. Immunol., 2005, Vol. 174, no. 9, pp. 5261-5269.

33. Kim M., Park H.J., Seol J.W., Jang J.Y., Cho Y.S., Kim K.R., Choi Y., Lydon J.P., Demayo F.J., Shibuya M., Ferrara N., Sung H.K., Nagy A., Alitalo K., Koh G.Y. VEGF-A regulated by progesterone governs uterine angiogenesis and vascular remodelling during pregnancy. EMBO Mol. Med., 2013, Vol. 5, no. 9, pp. 1415-1430.

34. Komatsu F., Kajiwara M. Relation of natural killer cell line NK-92-mediated cytolysis (NK-92-lysis) with the surface markers of major histocompatibility complex class I antigens, adhesion molecules, and Fas of target cells. Oncol. Res., 1998, Vol. 10, no. 10, pp. 483-489.

35. Korenevskii A.V., Milyutina Y.P., Zhdanova A.A., Pyatygina K.M., Sokolov D.I., Sel’kov S.A. Mass-Spectrometric analysis of proteome of microvesicles produced by NK-92 natural killer cells. Bull. Exp. Biol. Med., 2018, Vol. 165, no. 4, pp. 564-571.

36. Kowal J., Arras G., Colombo M., Jouve M., Morath J.P., Primdal-Bengtson B., Dingli F., Loew D., Tkach M., Thery C. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc. Natl. Acad. Sci. USA, 2016, Vol. 113, no. 8, pp. E968-E977.

37. Krueger A., Schmitz I., Baumann S., Krammer P.H., Kirchhoff S. Cellular FLICE-inhibitory protein splice variants inhibit different steps of caspase-8 activation at the CD95 death-inducing signaling complex. J. Biol. Chem., 2001, Vol. 276, no. 23, pp. 20633-20640.

38. Kumar S., Pan C.C., Bloodworth J.C., Nixon A.B., Theuer C., Hoyt D.G., Lee N.Y. Antibody-directed coupling of endoglin and MMP-14 is a key mechanism for endoglin shedding and deregulation of TGF-beta signaling. Oncogene, 2014, Vol. 33, no. 30, pp. 3970-3979.

39. Lash G.E., Robson S.C., Bulmer J.N. Review: Functional role of uterine natural killer (uNK) cells in human early pregnancy decidua. Placenta, 2010, Vol. 31 Suppl., pp. S87-S92.

40. Leonard S., Murrant C., Tayade C., van den Heuvel M., Watering R., Croy B.A. Mechanisms regulating immune cell contributions to spiral artery modification - facts and hypotheses - a review. Placenta, 2006, Vol. 27, Suppl A, pp. S40-S46.

41. Li P., Kaslan M., Lee S.H., Yao J., Gao Z. Progress in Exosome Isolation Techniques. Theranostics, 2017, Vol. 7, no. 3, pp. 789-804.

42. Liang S., Zhang J., Wei H., Sun R., Tian Z. Differential roles of constitutively activated ERK1/2 and NF-kappa B in cytotoxicity and proliferation by human NK cell lines. Int. Immunopharmacol., 2005, Vol. 5, no. 5, pp. 839-848.

43. Liang Y.J., Yang W.X. Kinesins in MAPK cascade: How kinesin motors are involved in the MAPK pathway? Gene, 2019, Vol. 684, pp. 1-9.

44. Lieberman J. The ABCs of granule-mediated cytotoxicity: new weapons in the arsenal. Nat. Rev. Immunol., 2003, Vol. 3, no. 5, pp. 361-370.

45. Liu K., He B., Xu J., Li Y., Guo C., Cai Q., Wang S. miR-483-5p targets MKNK1 to suppress Wilms’ tumor cell proliferation and apoptosis in vitro and in vivo. Med. Sci. Monit., 2019, Vol. 25, pp. 1459-1468.

46. Liu S., Yu D., Xu Z.P., Riordan J.F., Hu G.F. Angiogenin activates Erk1/2 in human umbilical vein endothelial cells. Biochem. Biophys. Res. Commun., 2001, Vol. 287, no. 1, pp. 305-310.

47. Lugini L., Cecchetti S., Huber V., Luciani F., Macchia G., Spadaro F., Paris L., Abalsamo L., Colone M., Molinari A., Podo F., Rivoltini L., Ramoni C., Fais S. Immune surveillance properties of human NK cell-derived exosomes. J. Immunol., 2012, Vol. 189, no. 6, pp. 2833-2842.

48. Male V, Sharkey A., Masters L., Kennedy P.R., Farrell L.E., Moffett A. The effect of pregnancy on the uterine NK cell KIR repertoire. Eur. J. Immunol., 2011, Vol. 41, no. 10, pp. 3017-3027.

49. Mandal A.,Viswanathan C. Natural killer cells: In health and disease. Hematol. Oncol. Stem Cell Ther., 2015, Vol. 8, no. 2, pp. 47-55.

50. Markov A.S., Markova K.L., Sokolov D.I., Selkov S.A., MARKMIGRATION. 2019: Russia.

51. Martinez-Lostao L., de Miguel D., Al-Wasaby S., Gallego-Lleyda A., Anel A. Death ligands and granulysin: mechanisms of tumor cell death induction and therapeutic opportunities. Immunotherapy, 2015, Vol. 7, no. 8, pp. 883-882.

52. Micheau O., Thome M., Schneider P., Holler N., Tschopp J., Nicholson D.W, Briand C., Grutter M.G. The long form of FLIP is an activator of caspase-8 at the Fas death-inducing signaling complex. J. Biol. Chem., 2002, Vol. 277, no. 47, pp. 45162-45171.

53. Mikhailova V.A. B.K.L., Vyazmina L.P., Sheveleva A.R., Selkov S.A., Sokolov D.I. Evaluation of microvesicles formed by natural killer (NK) cells using flow cytometry. Medical Immunology (Russia), 2018, Vol. 20, no. 2, pp. 251-254. doi: 10.15789/1563-0625-2018-2-251-254.

54. Mikhailova V.A., Ovchinnikova O.M., Zainulina M.S., Sokolov D.I., Sel’kov S.A. Detection of microparticles of leukocytic origin in the peripheral blood in normal pregnancy and preeclampsia. Bull. Exp. Biol. Med., 2014, Vol. 157, no. 6, pp. 751-756.

55. Murphy K., Weaver C. Janeway’s Immunology. Garland Science, Taylor & Francis Group, 2017. 924 p.

56. Naruse K., Lash G.E., Bulmer J.N., Innes B.A., Otun H.A., Searle R.F., Robson S.C. The urokinase plasminogen activator (uPA) system in uterine natural killer cells in the placental bed during early pregnancy. Placenta, 2009, Vol. 30, no. 5, pp. 398-404.

57. Okada H., Nakajima T., Sanezumi M., Ikuta A., Yasuda K., Kanzaki H. Progesterone enhances interleukin-15 production in human endometrial stromal cells in vitro. J. Clin. Endocrinol. Metab., 2000, Vol. 85, no. 12,pp. 4765-4770.

58. Osinska I., Popko K., Demkow U. Perforin: an important player in immune response. Cent. Eur. J. Immunol., 2014, Vol. 39, no. 1, pp. 109-115.

59. Philpott N.J., Scopes J., Marsh J.C., Gordon-Smith E.C., Gibson F.M. Increased apoptosis in aplastic anemia bone marrow progenitor cells: possible pathophysiologic significance. Exp. Hematol., 1995, Vol. 23, no. 14, pp. 1642-1648.

60. Pinto-Diez C., Garcia-Recio E.M., Perez-Morgado M.I., Garcia-Hernandez M., Sanz-Criado L., Sacristan S., Toledo-Lobo M.V., Perez-Mies B., Esteban-Rodriguez I., Pascual A., Garcia-Villanueva M., Martinez-Janez N., Gonzalez V.M., Martin M.E. Increased expression of MNK1b, the spliced isoform of MNK1, predicts poor prognosis and is associated with triple-negative breast cancer. Oncotarget, 2018, Vol. 9, no. 17, pp. 13501-13516.

61. Raposo G., Stoorvogel W Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol., 2013, Vol. 200, no. 4, pp. 373-383.

62. Riesbeck K., Billstrom A., Tordsson J., Brodin T., Kristensson K., Dohlsten M. Endothelial cells expressing an inflammatory phenotype are lysed by superantigen-targeted cytotoxic T cells. Clin. Diagn. Lab. Immunol., 1998, Vol. 5, no. 5, pp. 675-682.

63. Robson A., Harris L.K., Innes B.A., Lash G.E., Aljunaidy M.M., Aplin J.D., Baker P.N., Robson S.C., Bulmer J.N. Uterine natural killer cells initiate spiral artery remodeling in human pregnancy. FASEB J., 2012, Vol. 26, no. 12, pp. 4876-4885.

64. Schuler M., Green D.R. Mechanisms of p53-dependent apoptosis. Biochem. Soc. Trans., 2001, Vol. 29, Pt 6, pp. 684-688.

65. Sedgwick A.E., d’Souza-Schorey C. The biology of extracellular microvesicles. Traffic, 2018, Vol. 19, no. 5, pp. 319-327.

66. Si Y., Chu H., Zhu W, Xiao T., Shen X., Fu Y., Xu R., Jiang H. Concentration-dependent effects of rapamycin on proliferation, migration and apoptosis of endothelial cells in human venous malformation. Exp. Ther. Med., 2018, Vol. 16, no. 6, pp. 4595-4601.

67. Simak J., Gelderman M.P., Yu H., Wright V., Baird A.E. Circulating endothelial microparticles in acute ischemic stroke: a link to severity, lesion volume and outcome. J. Thromb. Haemost., 2006, Vol. 4, no. 6, pp. 1296-1302.

68. Singh R., Letai A., Sarosiek K. Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nat. Rev. Mol. Cell. Biol., 2019, Vol. 20, no. 3, pp. 175-193.

69. Smith S.D., Dunk C.E., Aplin J.D., Harris L.K., Jones R.L. Evidence for immune cell involvement in decidual spiral arteriole remodeling in early human pregnancy. Am. J. Pathol., 2009, Vol. 174, no. 5, pp. 1959-1971.

70. Smulski C.R., Decossas M., Chekkat N., Beyrath J., Willen L., Guichard G., Lorenzetti R., Rizzi M., Eibel H., Schneider P., Fournel S. Hetero-oligomerization between the TNF receptor superfamily members CD40, Fas and TRAILR2 modulate CD40 signalling. Cell Death Dis., 2017, Vol. 8, no. 2, e2601. doi: 10.1038/cddis.2017.22.

71. Sokolov D.I., Ovchinnikova O.M., Korenkov D.A., Viknyanschuk A.N., Benken K.A., Onokhin K.V., Selkov S.A. Influence of peripheral blood microparticles of pregnant women with preeclampsia on the phenotype of monocytes. Transl. Res., 2016, Vol. 170, pp. 112-123.

72. Susanto O., Trapani J.A., Brasacchio D. Controversies in granzyme biology. Tissue Antigens, 2012, Vol. 80, no. 6, pp. 477-487.

73. Svensson K.J., Christianson H.C., Wittrup A., Bourseau-Guilmain E., Lindqvist E., Svensson L.M., Morgelin M., Belting M. Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1. J. Biol. Chem., 2013, Vol. 288, no. 24, pp. 17713-17724.

74. Thornhill M.H., Li J., Haskard D.O. Leucocyte endothelial cell adhesion: a study comparing human umbilical vein endothelial cells and the endothelial cell line EA-hy-926. Scand. J. Immunol., 1993, Vol. 38, no. 3, pp. 279-286.

75. van der Pol E., Coumans F.A., Grootemaat A.E., Gardiner C., Sargent I.L., Harrison P, Sturk A., van Leeuwen T.G., Nieuwland R. Particle size distribution of exosomes and microvesicles determined by transmission electron microscopy, flow cytometry, nanoparticle tracking analysis, and resistive pulse sensing. J. Thromb. Haemost., 2014, Vol. 12, no. 7, pp. 1182-1192

76. Vermeulen K., van Bockstaele D.R., Berneman Z.N. Apoptosis: mechanisms and relevance in cancer. Ann. Hematol., 2005, Vol. 84, no. 10, pp. 627-639.

77. Wallace A.E., Fraser R., Cartwright J.E. Extravillous trophoblast and decidual natural killer cells: a remodelling partnership. Hum. Reprod. Update, 2012, Vol. 18, no. 4, pp. 458-471.

78. Wang M., Su P The role of the Fas/FasL signaling pathway in environmental toxicant-induced testicular cell apoptosis: An update. Syst. Biol. Reprod. Med., 2018, Vol. 64, no. 2, pp. 93-102.

79. Waters W.R., Harkins K.R., Wannemuehler M.J. Five-color flow cytometric analysis of swine lymphocytes for detection of proliferation, apoptosis, viability, and phenotype. Cytometry, 2002, Vol. 48, no. 3, pp. 146-152.

80. Xu R., Greening D.W., Zhu H.J., Takahashi N., Simpson R.J. Extracellular vesicle isolation and characterization: toward clinical application. J. Clin. Invest., 2016, Vol. 126, no. 4, pp. 1152-1162.

81. Yao L., Sgadari C., Furuke K., Bloom E.T., Teruya-Feldstein J., Tosato G. Contribution of natural killer cells to inhibition of angiogenesis by interleukin-12. Blood, 1999, Vol. 93, no. 5, pp. 1612-1621.

82. Zhang C., Gao F., Teng F., Zhang M. Fas/FasL Complex promotes proliferation and migration of brain endothelial cells via FADD-FLIP-TRAF-NF-kappaB pathway. Cell Biochem. Biophys., 2015, Vol. 71, no. 3, pp. 1319-1323.