Interleukin-8 Level in Pregnant Women with Toxoplasmosis in Kirkuk City

A cross sectional study was carried out in Kirkuk city from 15 th of June 2018 to 15 th of December 2018. The study included 100 pregnant women and 50 healthy individuals. Their ages ranged between 17-45 years old who were admitted to Kirkuk general hospital.. Molecular tests for real-time PCR and serological testing for detection specific Toxoplasma gondii IgM and IgG and Interleukin-8 level by using ELISA technique was done for patients and control. The study showed that the highest rate of anti T. gondii IgM+ IgGantibodies (10%) was recorded among pregnant women compared with 8% in the control group, while 22% of pregnant women were IgM+IgG+ compared 6.5% of the healthy control group. The study revealed that 40.91% of pregnant women with positive ELISA was positive by PCR compared with 0% of patients with negative ELISA results. The study showed that the highest rate of T. gondii infection (diagnosed by PCR) were recorded among pregnant women at age group 27-36 year (22.55%) and the lowest rate was within the age group 17-26 year. The highest mean level of IL-8 recorded PCR +ve groups, in pregnant women (79.2 ±53.2 ng/ml) compared with PCR –ve groups. There was a highly significant differences of IL-8 between pregnant women and the control group. The study showed that the highest mean level of IL-8 (77.61±60.4 ng/ml), in pregnant at 2 nd trimester of pregnancy, followed by 3 rd trimester. This study was concluded that a highly elevation of IL-8 level was correlated Toxoplasma infection in pregnant women and real time PCR is golden method in diagnosis of toxoplasmosis.


Introduction:
Toxoplasmosis is a very common infection caused by the obligate intracellular protozoan parasite [1]. This parasite is called Toxoplasma gondii widely distributed around the world.
Toxoplasma gondii can be vertically transmitted to the fetus during pregnancy and may cause wide range of clinical manifestations in the offspring depending on the gestational age at which the primary maternal infection was acquired, the virulence of the parasite and the immunologic development of the fetus [2]. The women may have spontaneous abortions, stillbirths, or premature delivery in addition to various fetal anomalies [3]. The frequency of severe congenital

Results:
The study showed that the highest rate of anti T. gondii IgM+ IgG-antibodies (10%) was recorded among pregnant women compared with 8% in the control group, while 22% of pregnant women were IgM+IgG+ compared with 6% of the healthy control group as shown in Table 1. shown in Table 2. The Table 3 shows that the highest rate of those who had IgM+ and IgG -T. in pregnant women was positive by PCR (70%) and 50% of patients with IgM+ IgG+ antibodies with non-significant relation (P: >0.05). The present study showed that the maximum rate (30%) of pregnant women in third trimester of pregnancy were positive by PCR followed by 16.67 of second trimester pregnant women and 7.41 of first trimester pregnant women, Fig. 1. The study showed that the highest rate of T. gondii infection (diagnosed by PCR) were recorded among pregnant women within the age group 27-36 year (22.55) and the lowest rate was within the age group 17-26 year with non-significant relation (P: >0.05) as shown in Table 4. The study showed that the highest mean level of IL-8 was recorded in PCR +ve groups in pregnant women (79.2 ±53.2 ng/ml) compared with PCR -ve groups, Table 5. The study demonstrated that there was a highly significant differences in the level of IL-8 between pregnant women and the control group (P: 0.0001), Table 6. The study showed that the highest mean level of IL-8 (77.61±60.4 ng/ml), was recorded in pregnant women in the 2 nd trimester of pregnancy, followed by 3 rd trimester. The result was significant, Table 7.

Discussion:
Regarding The real-time PCR offered the possibility of assessing disease progression and treatment efficacy [1,2]. Menotti et al [13] found that T. gondii DNA was detected more accurately by PCR optimum sensitivity when compared with ELISA. Although serological testing has been one of the major diagnostic techniques for toxoplasmosis, it has many disadvantages, for example, it may fail to detect specific anti-Toxoplasma immunoglobulin G (IgG) or IgM during the active phase of T. gondii infection, because these antibodies may not be produced until after several weeks of parasitaemia [14]. Therefore the high risk of congenital toxoplasmosis of a fetus may be undetected because the pregnant mother might test negative during the active phase of T. gondii infection [15]. Nimri et al [13]  revealed that 49.48 % of seropositive Toxo-IgG was seen within 3 rd trimester of pregnancy with highly significant relation P<0.01. Al-Hussien et al [19] found that the highest infection rates were found at 26-30 age group, while the lowest infection rate were found at age groups 36-40. Al-Rawazq [8] found that the seropositivity was observed higher in the age group between 20 to 30 years (37.1%). Khalil [20] found that Toxoplasma antibodies increase with age especially in the age group 25-30 years. This association does not mean that older age is a risky factor to predisposed to infection but might be explained by the older the person, the longer time being exposed to the causatiev agent and may retain a steady level of anti-Toxoplasma IgM in serum [21].
The chemokine response is dependent on invasion by live tachyzoites and subsequent host cell lysis. IL-8 is responsible for activation and recirculation of neutrophils and neutrophils  [23] indicated an increase of IL-8 in patients compared with healthy control and the highest mean level was recorded within the age group 29-39 year. Ali et al [24] indicated that the mean serum concentration of IL-8 in chronic and acute phase of T. gondii infection in pregnant women were more elevated than in healthy control. The chemokine response is dependent on invasion by live tachyzoites and subsequent host cell lysis. Furthermore, supernatants or lysates from T.gondii infected fibroblasts could elicit significant IL-8 secretion [25].
Increased level of IL-8 correlates with early acute inflammation or with a reactive form of toxoplasmosis. IL-8 is responsible for activation and recirculation of neutrophils and neutrophils can phagocytose and kill or inhibit tachyzoites of Toxoplasma and showed that human intestinal epithelial cells infected with T. gondii elicit rapid secretion of IL-8, so it has an important role in innate immunity in response to Toxoplasma [26]. In agreement with the present results, Borges et al [27] found that IL-8 was significantly increased in acute with early acute inflammation or with a reactive from toxoplasmosis in pregnant women.

Conclusions:
It was concluded that T. gondii infection was a highly related to elevation of IL-8 level in pregnant women and real time PCR is golden method in diagnosis of toxoplasmosis