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View of Assessment Study of Chemokine L5 Level in Hepatitis B Virus Patient and Compared with Risk Groups at Al Najaf City

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Assessment Study of Chemokine L5 Level in Hepatitis B Virus Patient and Compared with Risk Groups at Al Najaf City

Hasanain Imad SadeqKashef Al-Ghetaa1* and Fahem M.A-Shebly2

1,2Department of microbiology, College of veterinary, University of Kufa, Najaf, Iraq.

*[email protected]

Abstract

Hepatitis B virus infection (HBV) is still a major global health problem, which is an infectious disease that attacks the liver and a affects the lives of 325 million people worldwide and causing 1.4 million deaths a year. This study was designed to investigate and detection the levels of CXCL5 in the serum of hepatitis B virus (HBV) patients and risk group disease that infection patients (thalassemia, cancer and renal failure) by using enzyme-linked immunosorbent assay (ELISA) technique and compared level of CXCL5 among them and with the of normal level at Al-Najaf city. According to the best of our knowledge, this study is the first to display the levels of CXCL5 in different disease patients at al Najaf city. A total 155 sample were use in this study, that collection from different hospitals in Najaf city. The study design was included two experiments;The first was to indicated the HBV patients to ensure the infection of patients by HBV in additionally to take a history from each patient of risk groups disease for categorizing them into thalassemia, cancer, and renal failure patients’ groups. However, the second experiment detected the levels of CXCL5 in the HBV, thalassemia, cancer, and renal failure patients in different regions of Najaf City. The result showed that no significant between the levels of HBV (87.11 ± 2.87) and healthy (63.11 ± 11.66) groups, in additionally, the result showed the increased in the levels of CXCL5 in Renal Failure (140.4 ± 6.4) , Cancer (178.2 ± 6.8), and Thalassemia (118.9 ± 6.0). On conclusions, the present study confirmed the levels of CXCL5 is increased in the patients suffered HBV, cancer, renal failure and thalassemia in different levels among the healthy man.

Introduction

Hepatitis B virus infection remains a health major problem at the global level, which is an infectious disease that attacks the liver and affects the lives of more than three hundred million people, and the number of deaths in it reaches nearly one million and four hundred thousand

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people worldwide. While effect on 15 million people only on the European Region in 2019 and estimated that3.5% of the population suffers from chronic HBV (B) infections in world as well showed at 2015 (World Health Organization, 2015;2019). The Scientific evidences reports explainedthat immunity have ability vivacious role in initiating, maintaining immunological, regulating homeostasis and inflammation in many physiological and pathological processes such as HBV infection (Cheng et al., 2017), that by produced cytokines and chemokines in different stage and various immune cell in body (Yoshio et al., 2018). Chemokines are secretions of innate immune cells belonging to families of small cytokines, furthermore that coordinate and management the leukocyte migration during immunological processes and inflammation (Leighton et al., 2018). Chemokines CXC is one of four types of Chemokines interacting with G protein and chemokine receptors in immune cell (Mélik and Rostène, 2008). Several chemokines of the CXC are involved in neutrophil chemotaxis, also is noteworthy that the epithelial neutrophil that activates peptide 78 (ENA-78) is a type of CXC called CXCL5, are important neutrophil chemoattractant and proangiogenic factors and act in the setting of innate immunity (Stefano et al., 2019). The major causes of Hepatocellular carcinoma and liver cirrhosis is chronic infection by HBV,as it is characterized by the presence of a direct association between cirrhosis of the first stage and infection with hepatitis B virus, with no clear or special symptoms (Kanda et al., 2019; Hu et al., 2019). It has been noted here that some complications such as variceal bleeding, encephalopathy, dropsy, and spontaneous bacterial peritonitis according to some researchers that mean the disease has developed into the incurable state (Salehi et al., 2019). As well as current studies fund that is interface between immune response in HBV infection and risk group disease. However, CXCL5, which has recently been shown to be participate in Thalassemia(Taghavifar et al., 2019), cancer (Li et al., 2011) and renal failure (Haider et al., 2019). Therefore, it is very necessary to make an early diagnosis of viral inflammatory liver fibrosis in the early stages and to distinguish between pathological risks diseases. In this study, CXCl5 was detected in serum level of hepatitis patients and risk group disease (thalassemia, cancer and renal failure) by using ELIZA technique and compared level of CXCl5 among them and with the of normal level.

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Materials and Methods Study Site

The study was approved by the Institutional Review Board of Faculty of Veterinary in the University of Kufa and the written informed consent was obtained from each participant. The study was carried out at the Al Sader General Hospital, Al Zahra Hospital, Kidney Disease Center in Najaf and Blood bank center in Najaf and the samples from the patient of HBV and risk gropes disease (thalassemia, cancer and renal failure patents) it was placed in the icebox containing ice before being brought over the laboratory in Research Laboratory in Department of Microbiology in Faculty of Veterinary in the University of Kufa.

Experiment design

The study design was included two experiments. The first was to indicated the HBV patients in the different hospitals in Najaf City by use the HBV kit to ensure the infection of patients by HBV and take a history from each patient of risk groups disease for categorizing them into thalassemia, cancer, and renal failure patients groups. However, the second experiment detected the levels ls of CXCL5 in the HBV, thalassemia, cancer, and renal failure patients in different regions of Najaf City. Finally evaluated the different levels of CXCl5 among the serum of HBV patients and risk group disease (thalassemia, cancer, and renal failure) with control.

Patient Specimens

After taken history from patients five milliliters of venous blood were collected from each participant at his/her first admission to the hospitals. To harvest cell free serum, the blood was drawn into a sterile tube without anticoagulant. After leaving the tube in standing position for 20 minutes, samples were centrifuges at 20°C, 1,500g for 10 minutes, and the supernatant serum was quickly removed and stored immediately at until analysis. A total 155 sample was collection from different hospitals in Najaf city included (Al Sader General Hospital, Al Zahra Hospital, Kidney Disease Center in Najaf and Blood bank center in Najaf). Generally, the samples taken from different patients groups included (40 sample was taken from thalassemia patients, 40 sample was taken from renal failure patients, 40 samples was taken from cancer patients, 35 samples was taken from HBV patients) as well as, has been avoiding the corrupted sample during experiments, finally 80 samples used from risk group patients as described in

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following (18 sample of thalassemia, 19 sample of renal failure, 22 samples of cancer, 21 samples of HBV). For control group, 16 samples was collected from healthy people.

Hepatitis B virus detection by (ELISA): -

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human HBsAg. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. The 35 samples was taking from patients to detection the HBV patients, finally 21 samples was used in experiment.

Epithelial-derived neutrophil-activating peptide 78 by (ELISA): -

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human ENA-78/CXCL5.

Results

level of CXCL5 in patients and healthy control

Figure (1) showed an overview of differences in the level CXCL5 among risk group diseases with healthy. In short, the result showed that significant different at p>0.05 between healthy group and risk group excepted the HBVI which showed that is Non-significant different when compared to healthy group.

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Discussion

The present study aim wasn’t limited to the chemokine L5 level determination in hepatitis B virus patient and other risk groups disease only but also how can differentiation and understand relationship between hepatitis B virus patient and other risk groups disease according to the level of CXCL5 in blood. As far as we know, this study is the first displaying the CXCL5 levels in different disease patient at al Najaf city. The present study has been conducted to study the different levels of CXCL5 in hepatitis B virus patient and other risk groups disease included (cancer patient, thalassemia, renal frailer) in the serum by using enzyme-linked immunosorbent assay technique. Therefore, the experimental design of this study was aimed at standardizing and checked the hepatitis B virus patient as a way to assure that patient infected by hepatitis B virus and evaluation the CXCL5. Serum samples have been collected from Al Sader General Hospital, Al Zahra Hospital, Kidney Disease Center in Najaf and Blood bank center in Najaf.The current study found that elevation in level of CXCL5 in all group’s patients comparative with healthy status.The CXCL5 elevation due because it is the CXC-type chemokine family proangiogenic subgroup member also It is produced following stimulation of cells with the inflammatory cytokines. Has been found through the new studies that there is a relationship between CXCL5 and carcinogenesis and cancer development, as well as the CXCL5 is a member of CXC chemokines with neutrophilic chemoattractant which has been implicated in the pathological angiogenesis (Ichimura et al., 2014; Tacke et al., 2011).The present data recorded that serum level of CXCL5 was increased different grades as showed in figure (4-9), that according to side effect of infection or disease which several studies indicate that Chemokines have important immune roles for both rectum and colon (Yildirim et al., 2018). The CXCL5 expression in most human body such as epithelial cells within colorectal mucosa (Fujimura et al., 2018), as well as, the presence of CXCL5 in the epithelial cells of the renal tubules is related to its rejection of the human renal allograft (Higurashi et al., 2009). Therefore, we find that CXCL5 and what it attracts from neutrophils and T cells can make it possible to cause or enhance diabetic kidney injuries (Yu et al. al., 2017). Through investigation of CXCL5, CXCL5 was diagnosed in several types of cancer, especially in patients with bladder cancer. In addition, lung oncogenes and tumor suppressor genes regulate CXCL5 expression (Wu et. Al., 2017). Experimental evidence has demonstrated that there is an anti-tumor effect by the CXCL5-equivalent antibody or by using a chemical inhibitor of CXCR2 (Wang. et al., 2018).Actually, in patient populations and

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cancers there is a novel serum prognostic marker it is CXCL5 inducer of cell proliferation, migration, and invasion. (Hu et al., 2018) as well as agreed our result in figure (4-9), which that significant increases and superiority level of CXCL5 among all other patients in different groups than followed by renal failure in the serums as showed in current studies which indicates that knowledge of CXCL5 levels can be used to indicate diabetic nephropathy (Higurashi et al., 2009;

Wu et al., 2021). Some studies have suggested that serum CXCL5 levels are not necessarily an early definitive diagnostic factor for the disease as it is in the diagnosis of colorectal cancer (Wu et al., 2017; Yildirim et al., 2018; Bustani and Baiee 2021). Current studies have suggested that it is possible to increase the effectiveness of immunotherapy and to reduce the speed of tumor progression by blocking the transmission of CXCL5 / CXCR2 signals (Zhang et al., 2021). The current study also indicated the necessity of investigating the exact environment of the tumor and the factors affecting it, as well as the precise immune environment of cancer because the feasible method for treating cancer immunologically depends on these environments (Wu et al., 2020).

In conclusions

On conclusions, the present study confirmed the levels of CXCL5 is increased in the patients suffered HBV,cancer, renal failure and thalassemia in different levels among the healthy man. In additionally the current surveys is proved that significant different in levels of the CXCL5 among the HBV,cancer, renal failure, thalassemia and healthy groups can used in initially detection of disease.

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