Aqueous cytokine levels associated with severity of type 1 retinopathy of prematurity and treatment response to ranibizumab

  • Jiao Lyu
  • Qi Zhang
  • Haiying Jin
  • Yu Xu
  • Chunli Chen
  • Xunda Ji
  • Xiang Zhang
  • Yuqing Rao
  • Peiquan ZhaoEmail author
Basic Science



To determine the aqueous humor levels of cytokines in eyes with type 1 retinopathy of prematurity (ROP) before primary intravitreal injection of ranibizumab (IVR).


Forty-nine infants with type 1 ROP (56 eyes of 28 infants in the threshold ROP group and 42 eyes of 21 infants in the type 1 pre-threshold ROP group) received primary IVR and 49 aqueous humor samples were obtained preoperatively. Aqueous humor samples from 15 infants (15 eyes) undergoing congenital cataract surgery were used as controls. The concentrations of 27 cytokines were measured by a multiplex bead assay. Infants with persistent, recurrent, or progressive ROP after IVR were retreated.


The preoperative aqueous levels of 16 cytokines were significantly different among type 1 pre-threshold, threshold ROP, and control groups (P < 0.05). The concentrations of vascular endothelial growth factor (VEGF) (P < 0.001), interferon-γ (P < 0.001), interleukin (IL)-10 (P < 0.001), and IL-12 (P < 0.001) were the highest in the threshold ROP group, less in the type 1 pre-threshold ROP group, and the lowest in the control group. Retreatment was given to 55% of infants with ROP within a 48-week follow-up period after primary IVR. Higher VEGF (hazard ratio [HR] = 1.001, P = 0.001) and macrophage inflammatory protein-1β (HR = 1.085, P = 0.022) levels were independently correlated with ROP retreatment.


Higher aqueous levels of VEGF and inflammatory cytokines were associated with more severe type 1 ROP and ROP retreatment after primary IVR.


Retinopathy of prematurity Cytokine Aqueous humor Ranibizumab 



The authors thank the National Natural Science Foundation Project of China (81470642, to Peiquan Zhao) for funding this study. The authors also thank the participating patients and the medical staff of Xinhua Hospital, School of Medicine, Shanghai JiaoTong University.


National Natural Science Foundation Project of China (81470642, to Peiquan Zhao) provided financial support in the form of research funding.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the Ethics Committee of the Xinhua Hospital (XHEC-D-2014-090) and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. 1.
    Blencowe H, Lawn JE, Vazquez T, Fielder A, Gilbert C (2013) Preterm-associated visual impairment and estimates of retinopathy of prematurity at regional and global levels for 2010. Pediatr Res 74(Suppl 1):35–49. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Flynn JT, Chan-Ling T (2006) Retinopathy of prematurity: two distinct mechanisms that underlie zone 1 and zone 2 disease. Am J Ophthalmol 142(1):46–59. CrossRefPubMedGoogle Scholar
  3. 3.
    Hartnett ME, Penn JS (2012) Mechanisms and management of retinopathy of prematurity. N Engl J Med 367(26):2515–2526. CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Early Treatment For Retinopathy Of Prematurity Cooperative G (2003) Revised indications for the treatment of retinopathy of prematurity: results of the early treatment for retinopathy of prematurity randomized trial. Arch Ophthalmol 121(12):1684–1694. CrossRefGoogle Scholar
  5. 5.
    Mintz-Hittner HA, Kennedy KA, Chuang AZ, Group B-RC (2011) Efficacy of intravitreal bevacizumab for stage 3+ retinopathy of prematurity. N Engl J Med 364(7):603–615. CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Huang Q, Zhang Q, Fei P, Xu Y, Lyu J, Ji X, Peng J, Li YA, Zhao P (2017) Ranibizumab injection as primary treatment in patients with retinopathy of prematurity: anatomic outcomes and influencing factors. Ophthalmology 124(8):1156–1164. CrossRefPubMedGoogle Scholar
  7. 7.
    Hu J, Blair MP, Shapiro MJ, Lichtenstein SJ, Galasso JM, Kapur R (2012) Reactivation of retinopathy of prematurity after bevacizumab injection. Arch Ophthalmol 130(8):1000–1006. CrossRefPubMedGoogle Scholar
  8. 8.
    Wong RK, Hubschman S, Tsui I (2015) Reactivation of retinopathy of prematurity after ranibizumab treatment. Retina 35(4):675–680. CrossRefPubMedGoogle Scholar
  9. 9.
    Tasman W (1988) Multicenter trial of cryotherapy for retinopathy of prematurity. Arch Ophthalmol 106(4):463–464CrossRefGoogle Scholar
  10. 10.
    Feng J, Qian J, Jiang Y, Zhao M, Liang J, Yin H, Chen Y, Yu W, Li X (2017) Efficacy of primary intravitreal ranibizumab for retinopathy of prematurity in China. Ophthalmology 124(3):408–409. CrossRefPubMedGoogle Scholar
  11. 11.
    Sato T, Kusaka S, Shimojo H, Fujikado T (2009) Simultaneous analyses of vitreous levels of 27 cytokines in eyes with retinopathy of prematurity. Ophthalmology 116(11):2165–2169. CrossRefPubMedGoogle Scholar
  12. 12.
    Sood BG, Madan A, Saha S, Schendel D, Thorsen P, Skogstrand K, Hougaard D, Shankaran S, Carlo W, Nnr n (2010) Perinatal systemic inflammatory response syndrome and retinopathy of prematurity. Pediatr Res 67(4):394–400. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Yu H, Yuan L, Zou Y, Peng L, Wang Y, Li T, Tang S (2014) Serum concentrations of cytokines in infants with retinopathy of prematurity. APMIS : Acta Pathologica, Microbiologica, et Immunologica Scandinavica 122(9):818–823. CrossRefPubMedGoogle Scholar
  14. 14.
    Kong L, Demny AB, Sajjad A, Bhatt AR, Devaraj S (2016) Assessment of plasma cytokine profile changes in bevacizumab-treated retinopathy of prematurity infants. Invest Ophthalmol Vis Sci 57(4):1649–1654. CrossRefPubMedGoogle Scholar
  15. 15.
    Shiraya T, Kato S, Araki F, Ueta T, Miyaji T, Yamaguchi T (2017) Aqueous cytokine levels are associated with reduced macular thickness after intravitreal ranibizumab for diabetic macular edema. PLoS One 12(3):e0174340. CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    International Committee for the Classification of Retinopathy of P (2005) The international classification of retinopathy of prematurity revisited. Arch Ophthalmol 123(7):991–999. CrossRefGoogle Scholar
  17. 17.
    Gopinathan G, Milagre C, Pearce OM, Reynolds LE, Hodivala-Dilke K, Leinster DA, Zhong H, Hollingsworth RE, Thompson R, Whiteford JR, Balkwill F (2015) Interleukin-6 stimulates defective angiogenesis. Cancer Res 75(15):3098–3107. CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Petreaca ML, Yao M, Liu Y, Defea K, Martins-Green M (2007) Transactivation of vascular endothelial growth factor receptor-2 by interleukin-8 (IL-8/CXCL8) is required for IL-8/CXCL8-induced endothelial permeability. Mol Biol Cell 18(12):5014–5023. CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Johnsen-Soriano S, Sancho-Tello M, Arnal E, Navea A, Cervera E, Bosch-Morell F, Miranda M, Javier Romero F (2010) IL-2 and IFN-gamma in the retina of diabetic rats. Graefe’s Archive Clinical Experimental Ophthalmology = Albrecht von Graefes Archiv fur klinische und Experimentelle Ophthalmologie 248(7):985–990. CrossRefGoogle Scholar
  20. 20.
    Agarwal M, He C, Siddiqui J, Wei JT, Macoska JA (2013) CCL11 (eotaxin-1): a new diagnostic serum marker for prostate cancer. Prostate 73(6):573–581. CrossRefPubMedGoogle Scholar
  21. 21.
    Feng J, Zheng X, Li B, Jiang Y (2017) Differences in aqueous concentrations of cytokines in paediatric and adult patients with Coats’ disease. Acta Ophthalmol 95(6):608–612. CrossRefPubMedGoogle Scholar
  22. 22.
    Behzadi P, Behzadi E, Ranjbar R (2016) IL-12 family cytokines: general characteristics, pathogenic microorganisms, receptors, and signalling pathways. Acta Microbiol Immunol Hung 63(1):1–25. CrossRefPubMedGoogle Scholar
  23. 23.
    Ma J, Mehta M, Lam G, Cyr D, Ng TF, Hirose T, Tawansy KA, Taylor AW, Lashkari K (2014) Influence of subretinal fluid in advanced stage retinopathy of prematurity on proangiogenic response and cell proliferation. Mol Vis 20:881–893PubMedPubMedCentralGoogle Scholar
  24. 24.
    Castellanos MA, Schwartz S, Garcia-Aguirre G, Quiroz-Mercado H (2013) Short-term outcome after intravitreal ranibizumab injections for the treatment of retinopathy of prematurity. Br J Ophthalmol 97(7):816–819. CrossRefPubMedGoogle Scholar
  25. 25.
    Wallace DK, Kraker RT, Freedman SF, Crouch ER, Hutchinson AK, Bhatt AR, Rogers DL, Yang MB, Haider KM, DK VV, Siatkowski RM, Dean TW, Beck RW, Repka MX, Smith LE, Good WV, Hartnett ME, Kong L, Holmes JM, Pediatric Eye Disease Investigator G (2017) Assessment of lower doses of intravitreous bevacizumab for retinopathy of prematurity: a phase 1 dosing study. JAMA Ophthalmology 135(6):654–656. CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Lorenz B, Stieger K, Jager M, Mais C, Stieger S, Andrassi-Darida M (2017) Retinal vascular development with 0.312 MG intravitreal bevacizumab to treat severe posterior retinopathy of prematurity: a longitudinal fluorescein angiographic study. Retina 37(1):97–111. CrossRefPubMedGoogle Scholar
  27. 27.
    Appelberg R (1992) Macrophage inflammatory proteins MIP-1 and MIP-2 are involved in T cell-mediated neutrophil recruitment. J Leukoc Biol 52(3):303–306CrossRefGoogle Scholar
  28. 28.
    Sato T, Kusaka S, Shimojo H, Fujikado T (2009) Vitreous levels of erythropoietin and vascular endothelial growth factor in eyes with retinopathy of prematurity. Ophthalmology 116(9):1599–1603. CrossRefPubMedGoogle Scholar
  29. 29.
    Velez-Montoya R, Fromow G, Guerrero N, Clapp MC, Quiros M, Rivera GA (2010) Intraocular and systemic levels of vascular endothelial growth factor in advanced cases of retinopathy of prematurity. Clin Ophthalmol:947.
  30. 30.
    Chen SN, Lian I, Hwang YC, Chen YH, Chang YC, Lee KH, Chuang CC, Wu WC (2015) Intravitreal anti-vascular endothelial growth factor treatment for retinopathy of prematurity: comparison between Ranibizumab and Bevacizumab. Retina 35(4):667–674. CrossRefPubMedGoogle Scholar
  31. 31.
    Ozaki H, Okamoto N, Ortega S, Chang M, Ozaki K, Sadda S, Vinores MA, Derevjanik N, Zack DJ, Basilico C, Campochiaro PA (1998) Basic fibroblast growth factor is neither necessary nor sufficient for the development of retinal neovascularization. Am J Pathol 153(3):757–765. CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Fang L, Barber AJ, Shenberger JS (2014) Regulation of fibroblast growth factor 2 expression in oxygen-induced retinopathy. Invest Ophthalmol Vis Sci 56(1):207–215. CrossRefPubMedGoogle Scholar
  33. 33.
    Legacy J, Hanea S, Theoret J, Smith PD (2013) Granulocyte macrophage colony-stimulating factor promotes regeneration of retinal ganglion cells in vitro through a mammalian target of rapamycin-dependent mechanism. J Neurosci Res 91(6):771–779. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jiao Lyu
    • 1
  • Qi Zhang
    • 1
  • Haiying Jin
    • 1
  • Yu Xu
    • 1
  • Chunli Chen
    • 2
  • Xunda Ji
    • 1
  • Xiang Zhang
    • 1
  • Yuqing Rao
    • 1
  • Peiquan Zhao
    • 1
    Email author
  1. 1.Department of Ophthalmology, Xinhua Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Department of OphthalmologyShengli Oilfield Central HospitalDongyingChina

Personalised recommendations