Detection of HPV-16 and HSV-2 among women with Chronic Cervicitis in Baghdad City

 

Areej Atiyah Hussein¹, Basim Mohammed Khashman²

¹University of Diyala, College of Medicine, Department of Microbiology, Iraq

²University of Baghdad, Iraqi National Cancer Research Center, Iraq

 

ABSTRACT:

Background: Cervicitis is inflammation of cervical, it’s associated with most commonly of sexually transmitted viruses such as human papilloma virus and herpes simplex virus. Objective: To estimate the fre­quency of infection with human papilloma virus -16 and herpes simplex virus-2 in women with cervicitis and related with age and severity in Baghdad city. Patients and Methods: Retrospective study was conducted in 50 formalin-fixed paraffin embedded tissues from women with chronic cervicitis were collected from the archives of the Department of Pathology, Teaching Laboratories in Baghdad Medical City during the period from February 2018 till April 2018. Immunohistochemical staining of human papilloma virus -16 and herpes simplex virus -2 were determined in study group. Results: The expression of human papilloma virus -16 and herpes simplex virus -2 in women tissues with chronic cervicitis was 40% (20 out of 50). Most positive viral cases 10(50%) and 12(60%) were recorded in age group (20-35) years respectively; non-significant differences was noticed between viral infection and age of patients, majority of women were CIN1 44(88%) while other in CIN2 6(12%) also non-significant differences was found between viral infection and stage of chronic cervicitis. Conclusion: Infection rate of human papilloma infection - 16 and herpes simplex infection - 2 in this investigation were lower than different reports from various nations, this could be connected with the social conduct of ladies around there, where they generally have just a single accomplice for the duration of their life.

 

 

 

INTRODUCTION:

Cervicitis is an inflammatory condition of the cervix that is generally considered to result from infection with a sexually acquired organism [1]. Cervicitis in women has many features in common with urethritis in men, death may occur [2]. Chlamydia trachomatis and Neisseria gonorrhoeae as the prime etiologic suspects of cervicitis. Different life forms incorporate herpes simplex infection (HSV), human papillomavirus (HPV) and Trichomonas vaginalis [3]. Vaginal release or intermenstrual draining are visit manifestations of cervicitis, in spite of the fact that it might be asymptomatic [4].

 

 

Human papillomaviruses is generally little and non-encompassed have a place with the Papillomaviridae family [5]. Human papilloma infections DNA is available in every single cervical carcinoma and in some cervicitis cases, with some land variety in viral subtypes [6]. To date, more than 100 kinds of HPVs have been represented, which are delegated generally safe and high-hazard composes as per their relationship with threatening tumors. High-danger HPVs encode two oncogenes, E6 and E7, which expect essential part in carcinogenesis [7].

 

All herpisviruses are fundamentally comparative, every ha an icosahedra center encompassed by a lipoprotein envelope. The genome is direct twofold stranded DNA, the virion doesn't contain a polymerase [8]. Herpes simplex infection 1(HSV-1) basically causes mouth, throat, face, eye, and focal sensory system diseases, while HSV2 fundamentally causes anogenital lesions [9]. Genital herpes simplex infection remains a typical viral sexually transmitted sickness, frequently subclinical and a noteworthy overall issue of ladies of conceptive age assemble [10]. Genital herpes simplex infection is common and involves in the cervix, vagina, and vulva. The recurrence of genital herpes has expanded significantly in the previous decades, especially in adolescents and young ladies. By the age of 40, around 20% of ladies are seropositive for antibodies against HSV-2 [11]. Herpes simplex infection cervicitis happens in 70-90% of essential HSV-2 diseases and around 70% of essential HSV-1 infection [12]. Anyway there are no indisputable information on seroprevalence of herpes simplex cervicitis in many parts of the world, this is halfway in light of the fact that the pervasiveness of this ailment is extremely hard to find out by case revealing, as the greater part of the cases are clinically in obvious [13].

 

Cervical intraepithelial neoplasia (CIN), otherwise called cervical dysplasia, is the irregular development of cells on the surface of the cervix that could conceivably prompt cervical disease [14]. Infections of the cervix are normal in youthful sexually dynamic ladies [15]. Non-neoplastic diseases are predominantly inflammatory however there are few publications on the subject compared to neoplastic diseases of the cervix. Common causes include chemical irritations secondary to douching or local trauma produced by pessaries and intrauterine contraceptive devices [16]. The present study aims to estimate the fre­quency of infection with human papilloma virus -16 and herpes simplex virus-2 in women with cervicitis and related with age and severity in Baghdad city.

 

Patients and Methods:

Fifteen formalin-fixed paraffin embedded tissues from women with chronic  cervicitis were acquired from the documents of the Department of Pathology, Teaching Laboratories in Baghdad Medical City, from February 2018 to April 2018 for review think about. The conclusion of these tissue blocks were principally in view of the acquired histopathological records of cervix biopsy tests that had went with in the doctor's facility research center. Corroborative histopathological re-assessment of each acquired tissue blocks was finished by authority pathologist. Age of women range 20-63 years, 44 patients had CIN1 and 6 patients had CIN1, classification done depending on site of the injury and numerous factors [14].

 

According to immunohistochemical staining manufacturer's protocol for the detection of anti-HPV-16 (E2 antibody [TVG261] Cat. No. ab 17185- Cambridge Science Park - England) and Anti-HSV-1+HSV-2 antibody (Cat. No. ab227317 - Cambridge Science Park - England). After deparaffinization and rehydration by use xylene, serially graded alcohol for 3 minutes and finally by distill water, all slides were treated for 10 minutes with 3% hydrogen peroxide to inhibit endogenous peroxidase activity then washed in phosphate-buffered saline. The slides were then incubated for 5 minutes at 37°C with protein block trailed by wash with PBS, then treated at dilution (1/100) of primary antibody  for HPV-16 and HSV-2 and incubated at 37°C in humidity chamber for one hour followed by rinsed gently with PBS. secondary antibody  were placed on tissue section at room temperature for 10 minutes, trailed by the expansion of streptavidine-HRP antibodies for 10 minutes at 37˚C. In the wake of washing, weakened fluid DAB was utilized to recolored the slides for 15-45 minutes at room temperature then counterstained with hematoxylin for 30 second, last washed well done by utilize running faucet water, at that point got dried out and mounting with DPX and the last was checked by review first at 10 then at 400 amplification.

 

Statistical Analysis:

All data presented in this paper was analyzed by use Chi-square test, P value < 0.05 was regarded as significant.

 

RESULTS:

The result of this study demonstrated that 20 (40%) of cervical tissues were positive for human papilloma virus-16 and herpes simplex virus- 2, statistical analysis were demonstrated significant differences between viral expression among women with chronic cervicitis while no significant differences was noticed between two viruses as shown in Table (1) and Figure 1. On the other hand among positive cases for viral infection 10 (50%) had co-infection (both viruses) as shown in Figure 2.

 

Table (1): Positive and negative immunohistochemistry expression of human herpes virus-2 and human papilloma virus-16 in studied group.

IHC results	Positive No (%)	Negative No (%)	Comparison of Significance P-value
Human papilloma virus -16	20 (40%)	30(60%)	1.00 Non- significant at p < .05
Human herpes virus-2	20 (40%)	30(60%)

 

Figure (1): A- HPV-16 positive expression of EBV, B- HSV-2 positive expression by immunohistochemistry in cervical tissues of women with chronic cervicitis section (Magnification power, 400).

 

Figure (2): Viral co-infection in women with cervicitis.

 

The age of women with chronic cervicitis in current study ranged from 20-63 years with mean age 36.84?? years; majority of infected women 24 (48%) had been in observed amongst age group 36-50 years followed by 20-35 years, non-significant correlation had noticed between viral infection and age of participant, high percentage of participants with CIN1 44(88%),while 6 (12%) were recorded within CIN2. There was non-significant differences (P<0.05) had noticed between viral infection and stage of chronic cervicitis as shown in Table (2).

Table (2): Immunohistochemical results of HPV-16 and HSV-2 as associated to age of patients and stage of disease in the studied group.

Variables	Total No (%)	Positive HPV-16 No (%)	Positive HSV-2 No (%)	Comparison of Significance P-value
Age groups 20-35 years 36-50 years 51-65 years Total	20 (40%) 24 (48%) 6 (12%) 50(100%)	10(50%) 8(40%) 2(10% 20(100%))	12(60%) 6(30%) 2(10%) 20(100%)	0.4675*
Stage of disease CIN1 CIN2 CIN3 Total	44(88 %) 6(12 %) 0(0 %) 50(100%)	14(70%) 6(30%) 0 20(100%)	16(80%) 4(20%) 0 20(100%)	1.00*

 *Non-significant at p < .05

 

DISCUSSION:

The results of present study had showed that HPV-16 and HSV-2 were over expressed in cervical tissues from women with chronic cervicitis. These results lower than reported by Mirzaie-Kashani et al., (2014) found that 98.7% (78 out of 79) of the tested samples of chronic cervicitis were positive for human papilloma virus [6]. Also this result agreed with the study done by [17][18]. Another examination discovered a few gatherings of women have been observed to be at a higher danger of creating CIN through sexually transmitted human papillomavirus particularly the high-risk HPV composes 16 or 18 [14]. Then again the rate of HSV-2 infection in women with incessant cervicitis cases was 20 out of 50 (40%), which is considerably higher than that detailed by past investigations discovered cervicitis is thought to happen in ∼15%-20% of women with clinically obvious essential HSV-2 genital infection [19]. Also agreed with [10]. Herpes simplex infection 1 may likewise cause cervicitis; be that as it may, the signs are regularly less serious and more often than not happen just amid the essential genital disease [20]. Herpes cervicitis winds up imperative when seen in a lady of regenerative age gather due to the danger of neonatal contaminations coming about because of introduction to HSV in the genital tract amid conveyance [15]. As the same procedures were followed for all the tested cases, it is most unlikely that the positives scored in the cervicitis cases are due to cross contamina­tion so the etiology remains to be established. The similarity and differences between this result and the result of other studies could be related with age of participant, immune status, type of test used in diagnosis, number of sexual partners, sample size, viral load and studied area.

 

The result of this study found that half positive viral cases had co-infection, this outcome concurred with consider concentrating on gynecological diseases other than HPV as hazard determinants of ensuing cervical neoplasia, relationship with cervical HSV disease gave the most astounding and measurably most critical increment in chance [21].

 

According to age group, this study found a higher number of positive cases for HPV-16 and HSV-2 in age group (20-25) years. This result was in line with other reports indicating that the infections of the cervix are regular in youthful sexually dynamic ladies [15][16]. Women can develop CIN at any age, however women generally develop it between the ages of 25 to 35 [14].

 

This study it has been found that the higher percentage (44 cases) were notice in CIN 1, anyway most CIN precipitously relapse. Left untreated, around 70% of CIN-1 will relapse inside one year, and 90% will relapse inside two years [22]. Around half of CIN 2 will relapse inside 2 years without treatment to cervical carcinoma in situ (CIS) happens in roughly 11% of CIN1 and 22% of CIN2 [23]. Human papilloma infection disease is important for the advancement of CIN however not all with this contamination create cervical neoplasia [24]. Another investigation demonstrate that no all women tainted with HPV can create CIN or cervical growth, however those with a HPV contamination that does not leave without anyone else and keeps going more than 1 or 2 years have a higher risk of improvement of a higher grade of CIN [25].

 

In conclusions, infection rate of human papilloma infection - 16 and herpes simplex infection - 2 in this investigation were lower than different reports from various nations, this could be connected with the social conduct of ladies around there, where they generally have just a single accomplice for the duration of their life. Further study is warranted to clarify the role of other microorganism with large sample size. Educational programs to protect from this viruses should be included.

 

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Accepted on 18.05.2019           © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(9):4443-4446.