Centro de Investigación en Enfermedades Infecciosas ...€¦ · Centro de Investigación en Enfermedades Infecciosas, Instituto Nacional de Enfermedades Respiratorias Ismael Cosío

Centro de Investigaciones en Enfermedades Infecciosas Figure 1. Ocular OI were clinically assessed. A . CMVr retinitis; B ., ocular syphilis with severe vitritis (seric VDRL+). A B RESULTS Opportunistic ocular infections (OI) are not uncommon in HIV infection. Ocular immune response in the aqueous humor (AqH) has many unique properties that are still under research. Understanding these patterns could lead to improved or novel treatments of ocular OI. One previous study 1 found a predominantly Th1 response in AqH of CMVr. Using a multiplex technology, we measured multiple cytokine and chemokine profiles in AqH and plasma of HIV-infected individuals with OI, before and after specific treatment. Cytokine Proﬁle in Aqueous Humor of HIV Paents with Ocular Opportunisc Infecons Malde Ruiz-Cruz, MD, MSc; Sanago Ávila-Rios, PhD; Christopher E. Ormsby, MSc; Claudia Alvarado-de la Barrera, PhD; Yuria Ablanedo-Terrazas, MD, MSc; Gustavo Reyes-Terán*, MD, MSc. Centro de Invesgación en Enfermedades Infecciosas, Instuto Nacional de Enfermedades Respiratorias Ismael Cosío Villegas, Mexico City, Mexico. MATERIAL AND METHODS We included HIV+ patients with the following OI: 23 with cytomegalovirus retinitis (CMVr), 16 with ocular syphilis and 7 with other OI. As control measurements we analyzed plasma from 15 healthy donors and AqH from 16 patients undergoing cataract surgery. Concentrations of 27 cytokines in AqH and plasma before and after OI treatment were assessed using a Luminex assay. Cytokine concentrations in AqH and plasma were compared between OI and HIV-infected individuals vs. controls, by Wilcoxon tests corrected for multiple comparisons, Eotaxin IL-15 IL-7 G-CSF IL-17 IL-8 GM-CSF IL-1α IP-10 IFN-2α IL-1β MCP-1 IFN-γ IL-2 MIP-1α IL-10 IL-3 MIP-1β IL-12p40 IL-4 RANTES IL-12p70 IL-5 TNF-α IL-13 IL-6 TNF-β Table 1. Cytokines and chemokines included in the Luminex panel. Th1 cytokine , Th2 , Th1/2 , Th17 , stim factor , chemokines , proinflammatory , antiviral . Figure 2. Cytokine and chemokine levels pre-treatment were increased significantly in AqH and plasma, except for plasma IL-12p70 and IL-17, which were decreased. Levels of GM-CSF, IL-1 α and IP-10 in were significantly higher in CMVr than syphilis patients. C: Control, Cv: CMVr, S: Syphilis, O:Other. All cytokines and chemokines shown have p<0.05, q<0.2 between HIV+ and controls, others are not shown. Asterisk p<0.05, q<0.2 between CMVr and syphilis. Concentration ( pg/μl) 0 600 C Cv SO G-CSF 0 15000 +178,880 +178,880 C Cv SO IFN- γ +315 0 200 C Cv SO 0 200 C Cv SO 0 60 +632 +514 C Cv SO IL-10 0 200 +324 +431 C Cv SO 0 350 C Cv SO IL-1α 0 1500 C Cv SO 0 8000 C Cv SO IP-10 0 35000 C Cv SO 0 700 +1,196 +1,218 C Cv SO MCP-1 0 12000 C Cv SO TNF- α +105.7 0 70 C Cv SO 0 100 C Cv SO Plasma & AqH * * RANTES 0 15000 C Cv SO IL-12p70 +388 0 250 C Cv SO IL-17 +839 0 250 C Cv SO Eotaxin 0 120 C Cv SO GM-CSF 0 120 C Cv SO * IFN-2 α 0 400 C Cv SO IL-2 0 50 C Cv SO IL-6 0 10000 C Cv SO IL-8 +3,889 0 2500 +5,677 C Cv SO MIP-1α 0 200 C Cv SO MIP-1β 0 400 C Cv SO Plasma only Aqueous humor only Plasma & AqH INTRODUCTION Figure 3. Several cytokine and chemokine levels decreased significantly in AqH after specific treatment, and significantly more in CMVr than syphilis in IFN-2 α and IFN- γ . Dot: before treatment, arrowhead: after treatment. 0 10 20 30 40 50 60 70 80 90 100 Dim.5 4.02% Dim.4 7.47% Dim.3 8.89% Dim.2 20.64% Dim.1 43.79% (IL - 12p70, TNFb ) ( TNFb ) Th1 Th2 Th1Th2 Th17 Pro CK SF AV hi lo hi lo hi lo hi lo hi lo hi lo hi lo hi lo Cumulative percentage of variance explained 0 5 10 15 -6 -4 -2 0 2 4 6 8 Dim.1 43.79% Dim.2 20.64% HIV Ctrl 0 5 10 15 -6 -2 0 2 4 6 8 Dim.1 43.79% Dim.2 20.64% -4 CMVr Syphilis Figure 4. Principal component analysis of cytokine patterns in AqH. HIV infection had an apparent distinct pattern than control ( A ), as well as CMVr and syphilis ( B ). The compo- sition of the dimensions ( C ) showed that Th1 and chemokines had the largest contributions. Ellipses have 95% confidence. CONCLUSIONS During CMVr and ocular syphilis infection as HIV OI, the AqH shows a distinct cytokine profile, largely independent of plasma levels. The pattern comprised many AqH cytokines and chemokines, suggesting that treatments aimed at specific ones might not be as effective as antimicrobial treatment. The pattern is mostly composed of Th1 and chemokine responses, in accordance to the previous study, and we additionally observed longitudinally that the levels decreased to near normal levels with specific treatment. 1 REFERENCE Iyer JV, Connolly J, Agrawal R, et al. Cytokine 2013; 64 :541-7 *[email protected] Eotaxin G-CSF GM-CSF IFN-2 α 120 0 15,000 0 0 250 IFN- γ IL-10 IL-1 α 0 500 CMVr Syphilis 0 400 0 1,500 50 0 IL-6 IL- 8 IP-10 MIP-1 α 0 2,000 0 800 0 30,000 0 150 MCP-1 MIP-1 β TNF -α 0 10,000 0 250 0 100 Concentration (pg/ μl) IL-2 * * CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis CMVr Syphilis 120 POSTER 848 A B C

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Centro de Investigación en Enfermedades Infecciosas ...€¦ · Centro de Investigación en Enfermedades Infecciosas, Instituto Nacional de Enfermedades Respiratorias Ismael Cosío