Abstract
Background:
Toxoplasma gondii, a protozoan parasite with a worldwide distribution, is considered to infect one-third of all humans. many species. The intracellular parasite Toxoplasma gondii causes toxoplasmosis. Numerous physiological abnormalities are documented in toxoplasmosis-infected women.
Objective:
This study aims to demonstrate the connection between cyclophilins, the phospholipase enzyme, and latent toxoplasmosis.
Methods:
The research was carried out between January 2022 and June 2022. out of 150 patients had blood samples drawn, 250 had serum samples drawn from women with toxoplasma gondi infection, and 50 had healthy samples drawn from Hila city, Iraq. To exclude subjects who had any medical disorders, information from the subjects was gathered via an interviewer-managed questionnaire. ELISA was used to examine the serum. Results: About 250 samples from women with infertility were infected with Toxoplasma gondii overall (24%) Enzyme-Linked Immunosorbent Assay was utilized to evaluate the levels of phospholipase and cyclophilin, while automated VIDAS family instruments were employed to determine the qualitative and quantitative anti-Toxoplasma-IgG-tests (ELISA). Since there was a substantial difference in the statistical analysis and a significant difference in the cyclophilin protein, parasite infection changed the quantity of the enzyme phospholipase.
Conclusion:
This study put forth the theory that toxoplasmosis infection. Our investigation showed that patients with toxoplasma Gondi infection had higher levels of cyclophilins and phospholipase than control subjects.
Keywords: Toxoplasma gondii, Phospholipase, Cyclophilin
1. BACKGROUND
Toxoplasma gondii, a protozoan parasite with a worldwide distribution, is considered to infect one-third of all humans. many species It can infect warm-blooded animals and is a significant zoonotic and veterinary illness due to its medical and veterinary importance as well as its potential as a model for cell biology and molecular investigations using anti-Toxoplasma-IgG. It is also a parasite that has been extensively researched. Most infected people are asymptomatic or only exhibit minor symptoms, but pregnancy-related T. gondii infection can result in severe brain impairment and even fetal mortality (1).
A coccidian parasite known as T. gondii alternates between sexually developing stages in felines and various asexual forms in a variety of intermediate hosts during its complicated and strictly regulated life cycle (2). Depending on the climate, oocysts can live for up to 18 months and are a major cause of infection for both humans and animals. Consuming meat from animals that have been consistently infected with T. gondii and have tissue cysts due to this infection undercooked or cured meat items is another way this infection is spread (3). Severe pathological symptoms of toxoplasmosis can include retinochoroiditis, myocarditis, and meningoencephalitis, all of which have the potential to be fatal. However, the majority of infected people show little to no physiological symptoms and are asymptomatic. Latent T. gondii infection was once believed to have little impact on public health due to its asymptomatic nature, except in situations where immunosuppression was also present (4).
It’s exciting to look into the connection between toxoplasmosis and cognitive disorders. It appears that toxoplasmosis is a complex parasitological illness; if a human serves as an “accidental intermediate host” and has a robust immune system, Toxoplasma spp (5).
In vertebrates and other organisms, the cyclophilin protein family is present. They attach to the anti-immunosuppressive drug cyclosporine, also known as cyclosporine A, which was originally employed to stop organ rejection. By catalyzing the trans-to-cis isomerization of peptide bonds on proline residues, these proteins’ peptidyl proline isomerase activity enhances protein folding (6). Human cells now contain 17 different types of cyclophilins, with cyclophilin A (CypA) being the most common and accounting for 0.1-0.6% of the total cytoplasmic protein. These cyclophilins have different structural properties (7). Cyclophilin A (CypA), which is significantly higher in the serum of preeclampsia (PE) patients, plays crucial functions in inflammation and oxidative stress. The most challenging kind of pregnancy-related hypertension disease is preeclampsia (PE) (8).
One of the most common infectious diseases with a global distribution is toxoplasma gondii infection. Around the world, congenital toxoplasmosis results in 1.20 million life years lost to impairment each year, however, it is frequently disregarded in many nations. T. gondii is linked to several neurological diseases in both mothers and their newborns, as well as abnormalities in the reproductive system (9).
Additionally essential to infection, some parasites’ life cycles and host immune control is CypA. They are potentially potential therapeutic targets, particularly for cyclosporine A, an immunosuppressant. Additionally, cyclosporine has been shown to have anti-parasitic properties in a variety of taxa, including several apicomplexans (10). Members of the phospholipase superfamily regulate lipid metabolism, membrane composition, cell signaling, and inflammation. thorough knowledge of the structure, traits, and biotechnological applications of phospholipase A2 and other phospholipases (11). The enzyme mammalian phospholipase D generates phosphatidic acid, a dynamic lipid secondary messenger involved in numerous cellular processes including metabolism, motility, and exocytosis (PLD) (12).
Phospholipase A2 greatly facilitates Toxoplasma gondii entrance into host cells (PLA2). They play a crucial role in the host cell’s reaction to the invasion of the parasite. The cellular phospholipids of PLA2 are hydrolyzed, generating several inflammatory lipidic mediators (13). These phospholipid-hydrolyzing esterases are significant virulence factors because they are essential for membrane dynamics during parasite invasion and egress from the host cell, as well as for replication and cell signaling.
2. OBJECTIVE
The purpose of this study, according to Flammersfeld et al. was to measure the levels of cyclophilin and phospholipase in toxoplasma Gondi-infected women after IgG and IgM detection (14).
3. MATERIALS AND METHODS
Population Study
The research was carried out between January 2022 and June 2022. 150 patients had blood samples drawn, 250 had serum samples drawn from women with toxoplasma gondi infection, and 50 had healthy samples drawn from Hila city. To exclude subjects who had any medical disorders, information from the subjects was gathered via an interviewer-managed questionnaire. ELISA was used to examine the serum. In ELISA, the serum is used to establish an antigen-antibody combination on microtiter plates coated with soluble antigens (assuming specific antibodies are present). The antigen-antibody combination is subsequently detected by the addition of a secondary enzyme-linked antibody unique to the host species (15).
Sample collection
Each woman’s venous blood was sampled for 5 milliliters (ml) under sterile conditions, placed in a gel tube, and allowed to warm up before 15 minutes of centrifugation at 1000 rpm. Simple tubes were used to collect the serum, which was stored at -20 C until the IgM, IgG, cyclophilins, and phospholipase concentrations were measured.
The immunological study by Enzyme-linked immunosorbent assay (ELISA):
Antibodies to T. gondii can be found using ELISA assays. ELISA was used to examine the serum. In ELISA, the soluble antigen is coated onto microtiter plates, and serum is then added to generate an antigen-antibody combination (if specific antibodies are present) A supplementary enzyme-linked antibody that is specific to the host species is added to help identify antigen-antibody complexes (15).
Statistical analysis
SPSS 23 statistical software was used to conduct all statistical analyses (SPSS Inc., Chicago, USA). Our results and data were presented in mean and standard deviation, along with comparison statistics between the healthy group and patients. The relationships among the categorical variables were examined using an Enova test. At level (p 0.05), significant variations are accepted.
Table 2. Effect of age on the concentration of PL and Cyclophilin.
| P value | M±SD Patients | Parameters | Age group |
|---|---|---|---|
| 0.000*** | 71.605±11.594 | PL | (20) |
| 22.961 ±2.246 | Cyclophyline |
4. RESULTS
Measurements of the blood levels of cyclophilin and phospholipase in the analyzed samples using the enzyme-linked immunosorbent assay (ELISA) are shown in Ttable 1 and Figure 1. When compared to controls, patients with Toxoplasma gondii had a significantly higher level of cyclophilin and phospholipase, according to the results of the t-test (P 0.05).
Table 1. Mean and standard division of PL and cyclophilin between patients and control.
| P value | M±SD | Parameter | |
|---|---|---|---|
| Control | Patients | ||
| 0.05* | 66.844±8.527 | 71.605±11.594 | PL |
| 0.02* | 20.023±2.878 | 22.961±2.246 | Cyclophilin |
Figure 1. Mean and standard division PL and cyclophilin between patients and control.
Table 1 shows the effect of age group on the concentration of cyclophilin and phospholipase The result showed a significant difference.
Figure 2 shows that the correlation between PL and cyclophilin was negative correlation and had no significance in the value of Cyclophilin.
Figure 2. The correlation between PL and cyclophilin was negative correlation and no significant value of R=–0.139 P= 0.54.
5. DISCUSSION
After the occupation of Iraq, toxoplasmosis infection rates increased to almost 40% from 2% in the 1980s (16). In all Iraqi governorates in 2016, 335 patients with toxoplasma infection were documented in Iraq (17). Different regions of Iraq have varying climatic conditions and cultural norms, which affect the prevalence of toxoplasmosis (18, 19). Infections caused by several bacteria often include phospholipase A2 (PLA2), which has been linked to host cell invasion and has a substantial pathogenic function. We have looked into the part PLA2 plays in the cellular invasion by the tachyzoite stage of the intracellular protozoan Toxoplasma gondii. This process involves multiple phases. We measured the preferential incorporation of 3H-uracil into developing intracellular parasites to assess T. gondii invasion of human fibroblast monolayers (20). Our research demonstrates that patients with toxoplasma infections have higher concentrations of phospholipase. This work supports a prior observation that phospholipase promotes Toxoplasma gondii host cell penetration (20). Here, we demonstrate that the supernatant of acoustically disrupted Calcium-dependent phospholipase A (PLA) activity is present in T. gondii. Fatty acids and lysolipids were found to be released when host cells were treated with fragments of disruption. gondii in a bioassay, fractions of acoustically disrupted T. gondii with PLA activity increased T. gondii host cell penetration.
This study’s raised cyclophilins concentration compared to the control study agrees with that of Ibrahim et al. (2009), which found that TgCyp18-induced NO generation was crucial in both inhibiting parasite replication and initiating the development of bradyzoites This gave rise to the theory that TgCyp18 can influence the immune response to protect its host from the highly dividing tachyzoite as well as to complete its life cycle by converting to the slowly replicating bradyzoite T. gondii would have highly developed systems for controlling and influencing host cell reactions by secreting TgCyp18 (21).
6. CONCLUSION
In conclusion, our investigation showed that patients with toxoplasma Gondi infection had higher levels of cyclophilins and phospholipase than control subjects.
Acknowledgement:
The authors would like to thank The Biology Department, College of Science, University of Babylon, and Al-Mustaqbal University College (grant number MUC-M-0222). are to be thanked by the authors for their assistance and for providing the facilities required for this work, and we thank Dr. Yasir Haider Al-Mawlah, Dr. Ameer Mezher Hadi DNA Research Center / University of Babylon for their kind support.
Author's contributions:
Conception and design of the study: Noor Abdul Redah Al-Kremy, Maher Ali Al-Qrashy. Drafting the manuscript: Maher Ali Al-Qrashy. Analysis and/or interpretation of data: Noor Abdul Redah al-kremy, Maher Ali Al-Qrashy. Final proofreading was made by the first author.
Conflict of interests:
The authors have declared no conflict of interest.
Financial support and sponsorship:
Nil
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