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دراسة مناعية وجزيئية لخمج القرنية بحمة الحلا - 1 (حمة الهربس البسيط - 1) == Immunological And Molecular Study Of Herpes Simplex Virus - 1 Keratitis
Author name:
نهاية كاظم سالم
Supervisor name:
عبد العظيم ياسين عبود البراك | فائز اسماعيل الشكرجي
General topic:
Medicine
Specific topic:
Microbiology - Immunology
Degree:
Master
University:
Mustansiriyah University
Language:
English
University location:
Baghdad
First pages:
19T1150 - p.pdf
Abstract:
يمثل التهاب القرنية (Keratitis) مشكلة كبيرة قد تؤدي الى فقدان البصر احيانا نتيجة لخدش القرنية او فقدان الوضوحية في القرنية بعد اختفاء الالتهاب, ويعتبر فايروس الحلا (الهربس) البسيط - 1Herpes Simplex Virus - 1(HSV - 1))) هو من اهم العوامل المسببة لمرض التهاب ال | Keratitis is a medical term referring to inflammation of the cornea. It is a serious problem, resulting in everlasting loss of vision as a secondary effect to corneal scarring or loss of clarity of the cornea after the inflammation has disappeared. Most common microbes causing viral corneal keratitis is Herpes Simplex Virus - 1 (HSV - 1). HSV - 1 is a double stranded DNA (dsDNA) virus belongs to the Herpesiviridae. It affects the majority of peoples and its infections have been frequently without clinical manifestation. Ocular HSV - 1 infections caused multiple pathologies with possibly the most damaging being herpetic stromal keratitis (HSK). This study aimed to evaluate the abilities of the immunological and molecular indicators to diagnose herpetic keratitis in the infected and contralateral eye of patients, as well as to assess the immune status of both eyes by using tears and corneal scraping. Fifty - two patients with herpetic keratitis involved in this study, they attended the outpatient clinics / Ibn - AL - Haithum Teaching Hospital from March 2014 until December 2014. For the purpose of comparison, thirty, apparently healthy age matched control subjects were also included. Tears were collected from infected eyes of the 52 patients and from 16 out of the 52 patients contralateral eyes as well as from 30 controls. Corneal scraping had been obtained from 16 patient's infected eyes with corneal ulcers (epithelial ulcers) out of the total patients (52) with herpetic keratitis. Indirect immunofluorescence assay technique (IFA) was applied to measure levels of immunoglobulins (sIgA and IgG) in tears specific to HSV - 1. Realtime - polymerase chain reaction RT - PCR test was used to diagnose HSV - 1 in corneal scraping and tears of patients and control subjects. Interlukin - 17 (IL - 17A), tumor necrosis factor - alpha (TNF - ?) and interferon - gamma (IFN - ?) were measured in tear samples from patients and control subjects by the ELISA sandwich method. In tear samples; results of RT - PCR showed that 39/52 (75%) of the total patient's infected eyes were positive, 12/16 contralateral eyes were positive (75%). Corneal scraping and tear samples were taken from 16 patients with corneal ulcer, tear samples 14/16 (87.5%) positive result, while the corneal scraping samples were 16/16 (100%) positive, this indicates clearly that the cornea scraping is more appropriate and sensitive sample to detect HSV - 1 than tear. All controls were RT - PCR negative. IFA results of IgA and IgG in tears of infected and contralateral eyes were found to be more than the concentration in tears of control eyes with a highly significant p - value (0.0001). Comparison between the infected and the contralateral eyes indicates that both patients' eyes expressed a significant humoral immune response against HSV - 1. Although there were high significant differences between both the mean value of tears IL - 17A in infected and contralateral eyes with control, while the comparison between patient both eyes gave non - significant difference. The mean of tears TNF - ? and IFN - ? concentration in infected eyes of patients as compared to control was found to be highly significant (p= 0.0001). In this study we compared between patients infected eye with control to evaluate the performance of different markers in tear as HSV - 1specific sIgA, HSV - 1 specific IgG, IL - 17, TNF - ? and IFN - ? by using statistical test called Receiver Operating Characteristic (ROC) curve, the sensitivity for all studied parameters ranged from 80% - 96%.At the same time the comparisons between patient contralateral eyes and controls also revealed a high sensitivity range for these parameters ranged from 81% - 100%.These findings proved the ability of these markers in confirming diagnosis of HSV - 1 keratitis. In conclusion, the high percentage of RT - PCR results for corneal scraping sample indicate that this sample and by using this technique is more appropriate and sensitive to detect HSV - 1 than tear samples and detect the shedding of virus in both eyes of the same patients. Both patient eyes indicated that they were a good source for HSV - 1 diagnosis. In spite of the high sensitivity of all studied markers and their diagnostic ability, TNF - ? showed a higher sensitivity. To assess the immunological status of Herpetic keratitis patients, the studied markers TNF - ?, IFN - ? and IL - 17 were evaluated and showed a highly significant differences which indicated there was an active inflammatory condition combined with HSV - 1 infection and finally it can be concluded that both humoral and cellular branches of immune responses were involved in this disease.
