Relationship between the serum level, polymorphism and gene expression of IL-33 in samples of recurrent miscarriage Iraqi women infected with toxoplasmosis.

One of the many warm-blooded hosts that toxoplasmosis-causing intracellular protozoan parasite Toxoplasma gondii can infect is humans. Cytokines are crucial to stimulate an effective immune response against T. gondii. Interleukin-33 (IL-33) is a unique anti-inflammatory cytokine that suppresses the immune response. The levels of cytokine gene expression are regulated by genetics, and the genetic polymorphisms of these cytokines play a functional role in this process. Single nucleotide polymorphisms (SNPs) are prognostic indicators of illnesses. This study aimed to determine whether toxoplasmosis interacts with serum levels of IL-33 and its SNP in miscarriage women as well as whether serum levels and IL-33 gene expression are related in toxoplasmosis-positive miscarriage women. Two hundred blood samples from patients and controls were collected from AL-Alawiya Maternity Teaching Hospital and AL-Yarmouk Teaching Hospital in Baghdad, Iraq from 2021 to 2022 in order to evaluate the serum level of IL-33 using ELISA test. For the SNP of IL-33, the allelic high-resolution approach was utilized, and real time-PCR was performed to assess gene expression. The results showed that compared to healthy and pregnant women, recurrent miscarriage with toxoplasmosis and recurrent miscarriage women had lower IL-33 concentrations. Additionally, there were significant differences among healthy women, pregnant women, and women with repeated miscarriage who experienced toxoplasmosis. Furthermore, no differences between patients and controls were revealed by gene expression data. The results revealed that recurrent miscarriage, pregnancy, and healthy women all had a slightly higher amount of the IL-33 gene fold. Additionally, the SNP of IL-33 data demonstrated that there was no significant genetic relationship between patients and controls. Recurrent miscarriage women with toxoplasmosis have showed significant differences from pregnant women in the genotypes GG and AA as well as the alleles A and G. There were notable variations between recurrent miscarriage with and without toxoplasmosis in terms of the genotypes AA and AC. The genotypes GG, AA, and allele A in recurrent miscarriage women with toxoplasmosis and recurrent miscarriage women is a protective factor. Taking together, there was a statistically significant negative correlation between toxoplasmosis and IL-33 gene expression, which calls for more quantitative investigation in order to fully comprehend the interaction of mRNA and protein.

The association of IL-3, IL-17A, and IL 27 serum levels with susceptibility to toxoplasmosis in recurrent abortion of Iraqi women.

Cytokines are a group of immunomodulatory proteins leading to a variety of immune reactions in the human; these cytokines play a significant role in the development of appropriate immune responses against T. gondii. This study aims to reveal the association of toxoplasmosis with serum levels of IL-3, IL-17A, and IL-27 in aborted women. The blood samples of patients and controls were collected from Al-Alawiya Maternity Teaching Hospital/Baghdad/Iraq from 2019 to 2020 for detecting anti-T. gondii antibodies (IgG and IgM) and the level of interleukins by ELISA. The results of TORCH by rapid test for recurrent abortion recorded 25.3% seropositive for anti-Toxoplasma antibodies, and 31.5% seropositive for one or more cases of TORCH test (Cytomegalovirus, Rubella, and Herpes). Whereas the results for anti-T. gondii IgG and IgM antibodies were shown elevated positivity percentages by ELISA test; these percentages were 56.2% in recurrent abortion women with significant differences (P < 0.05). The results suggested that the IL-3 serum concentration of pregnant women, recurrent abortion, and recurrent abortion with toxoplasmosis was declined versus healthy women with significant differences (p < 0.05). However, the results revealed that the concentration of IL-17A in recurrent abortion, and recurrent abortion with toxoplasmosis elevated versus healthy women and pregnant women with significant difference (p < 0.05). Whereas the results indicated that the IL-27 serum concentration elevated with significant differences in recurrent abortion with toxoplasmosis group compared to healthy women, pregnant women, and recurrent abortion. Interestingly, the serum levels for IL-27 increased comparing to the levels of IL-3 and IL-17A in all groups with significant differences (P < 0.05). In conclusion, it appeared in this study that the role of IL-3, IL-17A, and IL-27 in the maternal immune response during infections can lead to abortion.

Evaluation of Apoptosis, Proliferation, and Adhesion Molecule Expression in Trophoblastic Tissue of Women With Recurrent Spontaneous Abortion and Infected With Toxoplasma gondii.

Recurrent spontaneous abortion is an obstetric complication with undefined causes. Apoptosis, proliferation, and adhesion are considered important factors in the pathogenesis of abortion. This work aimed to determine Bax and Bcl-2 as a proapoptotic and antiapoptotic protein, Ki67 and P27kip as proliferative and antiproliferative proteins, and E-cadherin and CD44 as adhesion molecules in the trophoblastic tissues in cases with recurrent miscarriage. Immunohistochemistry and quantitative polymerase chain reaction analysis of Bax, Bcl-2, Ki67, P27kip, E-cadherin, and CD44 in paraffin-embedded sections of placental tissues obtained from 108 women were divided into 3 categories: 66 Toxoplasma gondii-positive women with recurrent abortion, 22 T. gondii-negative women with recurrent abortion, and 20 women with no history of abortion as a control group. The mean ratio of the expression of Bax and P27kip proteins was 35.3% and 36.1%, which is significantly higher than that of the second group (19.88 and 20.02%), and the third group (12.3% and 10.98%), while the mean ratio of the expression of Bcl-2, Ki67, E-cadherin, and CD44 proteins was 12.35%, 11.23%, 10.32%, and 9.97%, which is significantly lower than that of the second group (33.75%, 13.18%, 21.88%, and 23.29%) and that of the third group (38.58%, 39.27%, 37.98%, and 35.79%). The presence of proapoptotic protein (Bax) and antiproliferative protein (P27kip) at high levels and the presence of antiapoptotic protein (Bcl-2), proliferative protein (Ki67), and adhesion molecules (E-cadherin and CD44) in lower levels in the T. gondii-positive group clarify the mechanism involved in the induction of abortion and loss of pregnancy.

Seroprevalence and risk factors of Toxoplasma gondii infection in women with recurrent fetal loss from the province of Khyber Pakhtunkhwa, Pakistan.

BACKGROUND: In women with a bad obstetric history (BOH), infection is an established cause of recurrent fetal loss. A common infecting agent is the protozoan parasite Toxoplasma gondii (T. gondii). The aim of this study was to measure the prevalence of toxoplasmosis in women with recurrent fetal loss from the Khyber Pakhtunkhwa province of Pakistan. METHODS: The study included 360 females aged 16-40 years, of which 180 had a bad obstetric history (study group) and the other 180 had no such history (control group). Blood serum samples were tested for toxoplasma IgM antibodies by Enzyme Linked Immunosorbent Assay and for toxoplasma IgG antibodies using an Immunochromatographic technique. RESULTS: The overall seroprevalence of toxoplasma infection in study group females was 40.6% and in control group females it was 7.2%. Specifically, IgM prevalence was 12.8% in the study group and 1.1% in the control group. IgG prevalence was 23.9% in the study group and 6.1% in the control group. IgM and IgG combined prevalence was 3.9% in the study group cases. There is a statistically significant association between BOH and seropositivity for T. gondii (p < 0.0001, Chi square test). Various risk factors associated with T. gondii seroprevalence in study and control groups were analyzed. CONCLUSION: The seroprevalence of toxoplasmosis was significantly higher in women with a bad obstetric history compared to those with no such history. Associated risk factors had no significant effects on the results.

Unpasteurised milk consumption as a potential risk factor for toxoplasmosis in females with recurrent pregnancy loss.

In women with a bad obstetric history, certain infections are associated with recurrent foetal loss. One of the common infectious agents is a protozoan parasite, Toxoplasma gondii. The aim of this study was to assess unpasteurised milk consumption as a potential risk factor for toxoplasmosis in females with recurrent pregnancy loss from the province of Khyber Pakhtunkhwa, Pakistan. In this study, we recruited a total of 360 females, comprising a study group of 180 females with previous history of recurrent pregnancy loss and a control group of 180 females with no such history. Blood serum from the participants was analysed for Toxoplasma gondii IgM antibodies by enzyme linked immunosorbent assay. Among the study group, 23 (12.8%) females were serologically positive for IgM antibodies against Toxoplasma gondii, whilst 157 (87.2%) were IgM negative. In the control group, only two (4.8%) females were IgM positive, whilst 178 (95.2%) were IgM negative. Bad pregnancy outcome in the study group and control group was observed to be significantly different (p < .0001). In both of these groups, unpasteurised milk consumption was found as a major risk factor for Toxoplasma gondii infection. A routine serological investigation should be carried out in pregnant women to rule out toxoplasmosis and reduce the risk of recurrent pregnancy loss as well as congenital toxoplasmosis in newborns.Impact statementWhat is already known on this subject? Seropositivity for Toxoplasma gondii antibodies ranges from 7% to 51% in different regions of the world. The prevalence rate varies because of differences in climate, culture, food habits, behaviour, personal hygiene and cooking habits of different societies and ethnic groups. Various risk factors have been identified that contribute to a high prevalence rate of the disease, including consumption of raw or poorly cooked meat, physical contact with cats or cat litter, consumption of unwashed raw vegetables and fruits, drinking of contaminated water and milk. We presumed that consuming unpasteurised milk could be a potential risk factor for developing toxoplasmosis in pregnant women.What the results of this study add? This study demonstrates high seroprevalence of Toxoplasma gondii antibodies in females of child bearing age that have consumed unpasteurised milk and is a potential risk factor for developing toxoplasmosis.What the implications are of these findings for clinical practice and/or further research? Our findings suggest that primary preventive measures (personal hygiene, frequent hand washing and consuming pasteurised milk) should be taken by health surveillance authorities to focus on families, especially pregnant women, to educate them about personal hygiene, contact with cattle or using their milk and milk products. The latter is especially important to aware them about the hazards of consuming unpasteurised and contaminated milk.

Chronic Neospora caninum infection and repeat abortion in dairy cows: a 3-year study.

The serological status of Neospora was monitored in animals older than 6 months in a dairy herd with a 3-year history of prevalent N. caninum and N. caninum-associated abortions. The numbers of animals in the herd tested each year of the study period were 259, 222 and 231, respectively. A separate analysis was performed on the 122 animals persisting in the herd for the 3 years. The overall seroprevalence of N. caninum in the herd decreased from 31.7% in the first year to 24.8% in the second year and to 19.9% in the third year of the study, while the overall abortion rate decreased from 20.6% in the first year to 5.5% in the second year, and 9.9% in the third. These decreases occurred in response to control measures adopted from the second year onwards, such as culling Neospora-seropositive aborted animals and inseminating Neospora-seropositive dams with beef bull semen. Of the total number of abortions recorded in seropositive animals, 51% were repeat abortions that occurred in 36.8% of the animals with a previous history of abortion. The initial seroprevalence of Neospora in the 122 animals followed for the 3 years was 18%, increasing to 21.3% in the second and third years. Seroconversion only occurred in four animals during the second and third years of the study and abortion occurred only in seropositive individuals. Of the total number of abortions recorded in the 122 animals, 61.5% were repeat abortions that occurred in 26.7% of the animals with a previous history of abortion. These results indicate that Neospora seropositivity can be very stable through time and N. caninum infected cows can show a high rate of repeat abortions. The present data reinforce the idea that annual serological screening for Neospora can be an effective and rapid method of detecting N. caninum infection, such that control measures can be established at the farm level.

Repetitive abortion in Neospora-infected ewes.

Pregnant ewes inoculated with cultured Neospora caninum tachyzoites in 1995, 1996, or 1995 and 1996 aborted or delivered weak or clinically normal lambs in 1996. Nine of 11 ewes in the study had previously produced infected lambs or fetuses after being experimentally infected in 1995. Fetuses and lambs produced in 1996 showed histologic lesions and zoites indicative of Neospora. Serologic responses and production of infected fetuses/lambs indicated systemic neosporosis in the ewes during gestation, although tachyzoites could not be cultured from maternal tissues. The repetitive infection of fetuses, and resulting clinical and histopathologic findings in the present study are similar to those reported in naturally infected cattle, adding to the already established similarities of neosporosis between sheep and cattle.

Seroprevalence of toxoplasmosis in women with recurrent abortions/neonatal deaths and its treatment outcome.

Toxoplasmosis, a parasitic infection, varies in its prevalence in various countries. Some studies have suggested its role in the causation of abortions. We reviewed the records of Microbiology Department at Sher-i-Kashmir Institute of Medical Sciences, Srinagar (Kashmir) and found that out of 2371 women with recurrent abortions and 310 women with neonatal deaths tested for IgM antibody against toxoplasma, 1260 (53.14%) and 215 (69.35%) tested positive respectively. One hundred and twenty-two women with recurrent abortions and 55 women with neonatal deaths who had tested positive for IgM antibody were followed during subsequent pregnancy and were treated with spiramycin; 115 94.26%) in current abortion group and 35 (63.64%) in neonatal death group delivered normal babies. We discuss the role of seropositivity for toxoplasma in women during reproductive period.

Toxoplasmosis as a cause of repeated abortion.

The extent to which toxoplasmosis causes habitual abortion is still controversial. The present study was carried out on 100 cases of repeated abortions, and 40 multiparous pregnant women at 20th week gestation as a control. Tests for the presence of toxoplasma IgG and/or IgM antibodies were done for both groups. It was found that 19% of the selected cases, and 7.5% of the control were seropositive for IgM. The difference was statistically insignificant, therefore acute toxoplasmosis, most probably, is not related to habitual abortions, whereas 37% of the selected cases and only 10% of the control group were seropositive at high dilution for IgG antibody, this statistical significant difference indicates that chronic toxoplasmosis, most probably is a significant cause of repeated abortion. Also it has been found that toxoplasmosis has a significant relation to abortion at first trimester.