CXCL9 chemokine level is associated with spontaneous clearance and sustained virological response in Egyptian Chronic Hepatitis C patients receiving direct acting antivirals

Abstract

BACKGROUND:

Chronic Hepatitis C virus (HCV) infection is associated with progressive liver inflammation which in turn leads to cirrhosis and finally causes hepatocellular carcinoma (HCC). By different escape mechanisms, the virus succeeds to evade the innate and acquired immune responses to establish chronic infection.

AIM:

This study aimed to evaluate the level of chemokine CXCL9 and its correlation with some biochemical parameters in different subjects of HCV patients.

MATERIALS AND METHODS:

A total of 83 persons participated in this study including healthy subjects without both HCV antibodies and HCV RNA (22.9%), HCV treated responders accomplished SVR post treatment, with HCV antibodies and absence of HCV RNA (24.1%), spontaneous or natural clearance patients, with positive HCV antibodies and negative HCV RNA without treatment (26.5%) and chronic HCV-patients, with both positive HCV antibodies and HCV RNA with no treatment (26.5%). HCV RNA was quantitated by real time PCR and serum CXCL9 level was measured by ELISA commercial kit pre-coated with human MIG/CXCL9 antibody. Assessment of biochemical and hematological parameters was carried out.

RESULTS:

Data showed that, the level of CXCL9 was significantly increased in chronic individuals (627.1 pg/ml) ( $P<$ 0.001) than spontaneous clearance (107.76 pg/ml) and responder subjects (117.28 pg/ml) ( $P\leqslant$ 0.05). No correlation has been found between CXCL9 level and viral load. Furthermore, CXCL9 levels correlated variably with some biochemical and hematological parameters according to each subject.

CONCLUSION:

Serum Chemokine CXCL9 level is associated with spontaneous clearance of HCV and response to HCV treatment, which may be identified as a predictive marker among HCV patients.

Keywords

CXCL9 chemokine Chronic Hepatitis C virus spontaneous clearance

1. Introduction

Hepatitis C virus (HCV) is a predominant pathogen with high mortality rates all over the world [1]. HCV infection affects about 170 million individuals worldwide [2]. Although HCV infection rate is diminishing in the developing countries, however, it has been reported that over the next 20 years the mortality from liver diseases secondary to HCV will keep on rising [3]. Among individuals who are exposed to HCV, 50 to 80% developed chronic infection that may progress to cirrhosis and/or hepatocellular carcinoma (HCC) [4]. Chronic infection is established when HCV successfully evades the host immune responses [5].

Studying the immune response in individuals who spontaneously cleared HCV infection is crucial to the development of a possible prophylactic vaccine. The mechanisms leading to viral eradication are not fully understood [3]. It has been reported that, levels of interferon $\alpha$ / $\beta$ stimulated genes in the liver of acutely infected chimpanzees with HCV were elevated [6, 7], Furthermore, divergence and interference of innate immune pathways by viral proteins including structural and nonstructural ones support the role for innate immune responses in viral clearance [8]. Adaptive immune response in HCV infected patients triggers strong and specific CD4 ${}^{+}$ and CD8 ${}^{+}$ T-cell responses that appear in acute infection and could be associated with viral resolution [9]. About 15 to 50% of infected subjects spontaneously clear the virus [10]. Controlling HCV infection has been associated with studying the effect of host polymorphisms of chemokines, interleukins, interferon-stimulated genes, proteins and natural-killer-cell receptors [11]. During acute and chronic HCV infection, neutralizing antibodies are detected in patients’ sera [9]. By increasing the neutralizing antibodies titer, the opportunities of HCV natural clearance increases [12].

Cytokines are provided as protective or immune-pathological responses after HCV infection. CXCL9 is a small cytokine that belongs to the chemokine family of CXC. It is also known as MIG, Monokine induced by (IFN- $\gamma$ ) gamma interferon. CXCR3 acts as a receptor for CXCL9 that plays role in induction of chemotaxis, differentiation and multiplication of leukocytes leading to tissue extravasation. Thus, CXCL9 is particularly important in chronic HCV infection by endorsing the development of intrahepatic inflammation leading to fibrogenesis [13]. Immune reactivity occurs through recruitment of immune cells, such as cytotoxic lymphocytes (CTLs), natural killer (NK) cells, NKT cells, and macrophages. A study showed that intrahepatic and peripheral levels of CXCR3-associated chemokines are significantly elevated in patients with advanced necro-inflammatory activity and fibrosis [14]. The CXCL10 was significantly associated with lobular inflammation [15, 16]. It has been demonstrated that polymorphisms in CXCL9-11 genes were associated with liver fibrosis in chronic hepatitis C patients’ acute rejection of liver allograft [17, 18]. Inflammatory morphea is characterized by T helper type 1 cytokine disturbance; particularly CXCL9 expression was associated with disease activity in lesional macrophages. CXCL9 is a promising biomarker of disease activity in morphea [19]. Saraiva (2018) reported that serum levels of cytokines were increased in HCV-infected patients, CCL-2, CCL-3, CCL-4, CXCL-8, CXCL-10, IL-1 $\beta$ , IL-15, IFN- $\gamma$ , IL-4, IL 10, TGF- $\beta$ , FGFb, and PAI-1 decreased significantly similar to non-infected controls after antiviral therapy. Moreover, cytotoxic effector natural killer exhaustion and CD8 ${}^{+}$ T cells have vital role in the establishing persistent HCV infections [20]. The current study was conducted to determine the level of CXCL9 in cases with spontaneous HCV clearance and chronic HCV infected patients and its correlation with some biochemical and hematological parameters in healthy, responders, spontaneous and chronic HCV patients.

2. Patients and methods

This study has been conducted on healthy subjects (22.9%), responders (24.1%), spontaneous clearance (26.5%) and HCV chronic individuals (26.5%) (Fig. 1). Nineteen healthy individual (9 females and 10 males), with an average age 23–51 years served as healthy volunteers, with negative HCV antibodies and twenty individuals (7 females and 13 males), with an average age 33–60 years were identified as a responder for HCV treatment, responders’ patients had positive HCV antibodies before treatment and negative HCV RNA PCR post treatment. Twenty-two subjects (10 males and 12 females) were identified as spontaneous clearance group of patients (age 33–64), Spontaneous clearance patients had positive HCV antibodies and negative HCV RNA PCR without any treatment. Twenty-two subjects (8 males and 14 females) were identified as chronic HCV-infections (age 35–65), Chronic infection, patients had positive HCV antibodies and positive HCV RNA PCR. Samples from healthy, responders and spontaneous clearance individuals were selected from the village of Ameriyah and Al-Mahalla Al-Kubra, Gharbia. Samples of chronic HCV patients were obtained from the National Liver Institute in Shebin El-Kom. This study did not include non-responders’ cases after treatment (i.e. have both HCV antibodies and RNA) due to high successful percentages of success of treatment of Direct Acting Antivirals (DAAs) in Egypt [21].

Figure 1.

Distribution of the enrolled patients in the present study.

2.1 Antiviral regimen

In the responders’ individuals, they were treated with interferon only, which was injected weekly for a period of 48 weeks and most of them got a relapse and then took DAAs in the form of the triad treatment consisting of Sovaldi (Sofosbuvir), ribavirin and interferon. Only one patient got a relapse then took Harvoni (Ledipasvir $+$ Sofosbuvir) and had sustained virological response.

2.2 Blood sampling

The blood samples were collected from all individuals enrolled in this study and were divided into two parts; the first part was left to clot for 30 min to obtain the sera. The second part was kept with anti-coagulant agent (EDTA) for measuring the hematological parameters and collection of plasma samples. Plasma and sera were aspirated carefully and let to freeze at $-$ 20 ${}^{\circ}$ C till further investigations.

2.3 Biochemical and hematological analysis

Liver transaminases, aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) and albumin were measured by commercial kits supplied by SPINREACT, S.A.U. Ctra. Santa Coloma, Girona Spain. Serum total and direct Bilirubin were measured by spectrophotometer using kits (Diammond, Cairo, Egypt). HCV Ab was tested by rapid test device (ABON Biopharm, Hangzhou, Co., Ltd) and it is confirmed by HCV Ab Elisa (abia ${}^{\@setsize{\scriptsize}{8pt}{\viipt}{\@viipt}{\textregistered}}$ , Germany)

2.4 HCV-RNA quantification

HCV RNA was assessed by the Roche COBAS ${}^{\@setsize{\scriptsize}{8pt}{\viipt}{\@viipt}{\textregistered}}$ AmpliPrep/COBAS TaqMan ${}^{\@setsize{\scriptsize}{8pt}{\viipt}{\@viipt}{\textregistered}}$ HCV Quantitative Test v2.0 (Roche Molecular Systems Inc., Branchburg, NJ, USA). The lower detection limit was 15 IU/ml. HCV-RNA levels were assessed in all cohorts.

2.5 CXCL9 quantification

Serum CXCL9 level was measured in all cohorts by the commercially enzyme-linked immunosorbent assay according to manual procedures based on pre-coated plates with human MIG/CXCL9 antibody and using the Biotin double antibody sandwich technology to assay the Human monokine induced by interferon-gamma (MIG/CXCL9) BT Bioassay technology laboratory (Shanghai Korain Biotech Co., LTD, Cat.No E0049Hu). The minimum detectable level is 10.01 ng/L. The total count of red blood cells (RBCs), total count of white blood cells (WBCs), total count of platelets, hemoglobin and differential leucocytes were assessed by Hematological analyzer (Mindray CBC cell counter, India).

2.6 Statistical analysis

Data were analyzed using Graph Pad Instat software (San Diego, CA, USA). The data were expressed as mean $\pm$ SD. The significance of differences among treated groups and control were analyzed by means of one-way ANOVA followed by Tukey. Acceptable significance was considered when $P$ -value was $P\leqslant$ 0.05.

Table 1
The hematological parameters profile among cohort study

Parameters	Healthy		Spontaneous		Responders		Chronic		$P$ -value
Age (years)	37.47	$\pm$ 2.26	48.68	$\pm$ 2.41	48.85	$\pm$ 1.87	52.27	$\pm$ 2.05
Sex
(M)	10 (11.76%)		10 (11.76%)		13 (15.66%)		8 (9.64%)
(F)	9 (10.59%)		12 (14.12%)		7 (8.43%)		14 (16.87%)
Hb (g/L)	13.55	$\pm$ 0.3 ${}^{\text{a}}$	11.71	$\pm$ 0.28 ${}^{\text{b}}$	13.04	$\pm$ 0.43 ${}^{\text{a.b}}$	12.35	$\pm$ 0.4 ${}^{\text{a.b}}$	0.003 s.
RBCs (10 ${}^{6}$ /mm ${}^{3}$ )	4.66	$\pm$ 0.09 ${}^{\text{a.b}}$	4.55	$\pm$ 0.10 ${}^{\text{a.b}}$	4.87	$\pm$ 0.16 ${}^{\text{a}}$	4.34	$\pm$ 0.14 ${}^{\text{b}}$	0.031 s.
PLT (10 ${}^{3}$ /mm ${}^{3}$ )	236.89	$\pm$ 15.01 ${}^{\text{a}}$	224.1	$\pm$ 11.2 ${}^{\text{a.b}}$	203.55	$\pm$ 15.08 ${}^{\text{b}}$	271.1	$\pm$ 10.56 ${}^{\text{a}}$	0.003 s.
TLC(10 ${}^{3}$ /mm ${}^{3}$ )	6.13	$\pm$ 0.47 ${}^{\text{b}}$	6.47	$\pm$ 0.55 ${}^{\text{a.b}}$	6.62	$\pm$ 0.31 ${}^{\text{a.b}}$	8.02	$\pm$ 0.63 ${}^{\text{a}}$	0.055 s.
Neutrophils (%)	53.93	$\pm$ 2.98	50.35	$\pm$ 2.19	52.17	$\pm$ 2.57	51.49	$\pm$ 2.38	0.792 n.s.
Lymphocytes (%)	38.27	$\pm$ 2.80	41.50	$\pm$ 2.17	40.50	$\pm$ 2.71	40.51	$\pm$ 2.33	0.834 n.s.
Monocytes (%)	7.68	$\pm$ 0.37	8.15	$\pm$ 0.58	7.02	$\pm$ 0.30	8.01	$\pm$ 0.52	0.343 n.s.

All parametric values are expressed as means $\pm$ SE. $P$ -values lower than 0.05 were considered significant. Means that do not share a letter are significantly different. Where RBCs (10 ${}^{6}$ /mm ${}^{3}$ ): red blood cells: Male 4.5–5.6 & Female 3.8–5.0, Hb (g/L): Hemoglobin: Male 13.0–17.0 & Female 12.0–16.0, HCT (%): Male 40.0–52.0 & Female 36.0–42.0, TLC (10 ${}^{3}$ /mm ${}^{3}$ ): Total Leukocyte count: 4.0–11.0, Neutrophils (%): 40–60, Lymphocytes (%): 20–40, Monocytes (%): 2.0–8.0, PLT (10 ${}^{3}$ /mm ${}^{3}$ ): Platelets count: 150–400.

3. Results

3.1 The hematological profile among studied cohorts

Hematological analysis in the healthy, responder, spontaneous and chronic HCV subjects was assessed by determining red blood cells (RBCs), white blood cells (WBCs), hemoglobin (Hb), platelets and differential count of leucocytes. Results showed that there was a significant decrease in the hemoglobin level in the spontaneous clearance individuals as compared to healthy persons ( $P\leqslant$ 0.05). Furthermore, there was significant decrease in RBCs level in chronic subjects as compared to responder persons ( $P\leqslant$ 0.05), while no significant change in the level of RBCs between healthy and spontaneous subjects has been detected. Responder patients have also showed a significant decrease in the total number of platelets count when compared to chronic HCV persons ( $P=$ 0.003). Also, there was significant decrease in the TLC in healthy subjects compared to chronic ( $P=$ 0.05), while there was no significant change in the level of TLC between spontaneous clearance and responder subjects. Differential leucocytes were detected in all subjects and results showed that there was no change in the percentages of neutrophils and lymphocytes among different subjects (Table 1).

Figure 2.

Serum level of CXCL9 in healthy individuals, spontaneous, responders and chronic HCV patients as measured by ELISA.

Figure 3.

The correlation between CXCL9 serum level and HCV viral load. There is no correlation between serum level of CXCL9 and HCV viral load ( $r=$ 0.145), ( $P<$ 0.52).

Table 2

The biochemical parameters profile among cohort study

Parameters	Healthy		Spontaneous		Responders		Chronic		$P$ -value
GPT (U/L)	23.83	$\pm$ 2.48 ${}^{\text{b}}$	26.94	$\pm$ 3.05 ${}^{\text{b}}$	23.08	$\pm$ 2.06 ${}^{\text{b}}$	48.41	$\pm$ 41 ${}^{\text{a}}$	0.001 s.
GOT (U/L)	25.37	$\pm$ 1.74 ${}^{\text{b}}$	27.20	$\pm$ 2.59 ${}^{\text{a.b}}$	22.94	$\pm$ 1.17 ${}^{\text{b}}$	35.38	$\pm$ 3.72 ${}^{\text{a}}$	0.006 s.
Albumin (g/L)	4.84	$\pm$ 0.04 ${}^{\text{a}}$	4.49	$\pm$ 0.05 ${}^{\text{b}}$	4.47	$\pm$ 0.05 ${}^{\text{b}}$	4.59	$\pm$ 0.14 ${}^{\text{a.b}}$	0.012 s.
Direct bilirubin (mg/dl)	0.14	$\pm$ 0.01 ${}^{\text{b}}$	0.16	$\pm$ 0.01 ${}^{\text{a.b}}$	0.19	$\pm$ 0.02 ${}^{\text{a.b}}$	0.21	$\pm$ 0.02 ${}^{\text{a}}$	0.031 s.
Total bilirubin (mg/dl)	0.60	$\pm$ 0.02	0.67	$\pm$ 0.03	0.70	$\pm$ 0.06	0.66	$\pm$ 0.04	0.360 n.s.
Cxcl9 (pg/ml)	110.67	$\pm$ 16.80 ${}^{\text{b}}$	107.67	$\pm$ 19.82 ${}^{\text{b}}$	117.28	$\pm$ 31.93 ${}^{\text{b}}$	627.1	$\pm$ 169.8 ${}^{\text{a}}$	0.000 s.

All parametric values are expressed as means $\pm$ SE. $P$ -values lower than 0.05 were considered significant. Means that do not share a letter are significantly different. Where Direct Bilirubin (mg/dl): Up to 0.25, Total Bilirubin (mg/dl): Up to 1.0, Albumin (g/L): 3.5–5.2, Glutamate pyruvate transaminase: Up to 40, Glutamate Oxaloacetate transaminase: Up to 40.

Figure 4.

Correlation among CXCL9 level and liver function parameters. CXCL9 positively correlated with albumin, liver function GPT and GOT and negatively correlated with bilirubin (direct and total).

Figure 5.

The correlation among CXCL9 level and hematological parameters. CXCL9 negatively correlated with total number of RBCs, hemoglobin, total leukocytic count, neutrophils and monocytes and positively correlated with platelets and lymphocytes.

3.2 The biochemical parameters profile among studied cohorts

The activity of GPT and GOT were determined in all subjects under the study. Results showed that serum GOT activity had no significant difference in the serum of all subjects, while GPT activity was significantly increased in the serum of chronic HCV patients when compared with healthy patients ( $P\leqslant$ 0.0001) (Table 2). Albumin levels showed significant decrease in all responders, spontaneous clearance and chronic patients when compared to the healthy patients ( $P\leqslant$ 0.05) (Table 2). Furthermore, total and direct bilirubin in different subjects, showed that there was significant increase in the levels of direct bilirubin in chronic HCV as compared to the healthy individuals ( $P\leqslant$ 0.05).

3.3 Levels of CXCL9 in the different groups under the study

The serum mean levels of CXCL9 were significantly higher ( $P<$ 0.001) in chronic hepatitis individuals 627.1 pg/ml compared to healthy subjects 110.67 pg/ml and spontaneous clearance subjects 107.76 pg/ml. Also the CXCL9 level was significantly higher compared to responders 117.28 pg/ml (Fig. 2). Results showed that there was no correlation between the levels of CXCL-9 and HCV expression level by gene expression analysis (Fig. 3).

3.4 The correlation between CXCL9 and some biochemical parameters

Data showed that CXCL9 level in serum was positively correlated with albumin, liver function GPT and GOT and negatively correlated with bilirubin (direct and total) (Fig. 4).

Statistical analysis showed that there was negative correlation between the level of CXCL9 and the total number of RBCs, hemoglobin, total leukocytic count, neutrophils and monocytes and positive correlation between platelets and lymphocytes with CXCL9 (Fig. 5).

4. Discussion

The present study aimed to assess the association between CXCL9 level in different patients groups; namely acute, chronic HCV patients, DAAs- responders and infection clearance, as well as assessment of correlation CXCL9 level with some hematological and biochemical features. In general, a significant decrease in the hemoglobin level in the spontaneous clearance group was observed compared to healthy persons according to complete blood picture findings. Also, DAAs-responder patients group showed a significant decrease in the total number of platelets count when compared to chronic HCV persons in agreement with previous studies that reported a significant decrease in pretreatment platelet count in the sustained virological response (SVR) [22, 23].

A significant increment in CXCL9 level in chronic patients (627.1 pg/ml) compared with spontaneous clearance (107.76 pg/ml) and responders groups (117.28 pg/ml), which complies with the findings of Zeremski et al. [24], while no significant difference between CXCL9 level in healthy (110.67 pg/ml) and responder groups (117.28 pg/ml) after DAAs has been observed. These findings agree with [25] who reported that levels of CXCL9 declined during therapy in both responders and non-responders and among sustained responders, levels of CXCL9 remained low after completion of therapy while in non-responders, the drop in CXCL9 was transient. Also it was stated that CXCL9 levels were not significantly different in sustained responders after treatment when compared with healthy control subjects. Also, Butera et al. [25] has studied the relationship between serum levels of CXCR3-associated chemokines and response to antiviral therapy; and revealed that CXCL9 and CXCL10 were decreased in those for whom treatment had been successful. It is worth mention that there is no relapses/recurrence during DAAs treatment throughout the follow up period for the HCV infection patients using combination therapy of SOF/DCV that proved to be safe and effective in the treatment of chronic HCV genotype-4 infection the predominant type in Egypt [21]. A pervious study conducted by our team work on Egyptian patients also showed that CXCL10 plasma level was elevated in chronic patients and low CXCL10 level was associated with therapeutic response [26]. No correlation has been found between CXCL9 level and viral load, which could be attributed to the possibility that it does not directly contribute to antigen-specific T cells recruitment for viral replication control rather than a neutral or detrimental inflammatory reaction, and comes in agreement with Butera et al. [25] and Johansson et al. [27] findings. On the other hand, while levels of CXCL9, CXCL10, and CXCL11 chemokines have been observed to be significantly elevated in chronic HCV patients; levels of CXCL9 and CXCL10 were declined after successful antiviral therapy, and similarly levels of CXCL11 did not decline significantly during or in the first 6 months after therapy.

Upon monitoring the biochemical parameters in our cohorts, it has been observed that serum GPT was significantly increased in chronic HCV patients’ sera comparing to healthy persons, while albumin levels were significantly decreased among responders, spontaneous clearance and chronic patients’ groups when compared to the healthy subjects. This comes in line with previous studies that reported elevated serum GPT levels correlated with viral loads in chronic HCV patients, which may have clinical relevance [28, 29]. Moreover, our findings revealed that the degree of albumin synthesis capacity was lower in DAAs-responders and chronic HCV patients groups than spontaneous patients group, which complies with the findings of both [30, 31] who indicated that the hepatic capacity of albumin synthesis was affected by the degree of liver fibrosis. Total and direct bilirubin in the different subjects showed that there was a significant elevation in direct bilirubin levels in all groups under study comparing to healthy individuals in line with previous studies that reported an elevation of serum bilirubin levels during chronic HCV infection [32]. Finally, our results revealed a positive correlation between GPT and both CXCL9 ( $r=$ 0.335, $P<$ 0.001) and CXCL10 (IP-10) levels ( $r=$ 0.423, $P<$ 0.001) serum levels, which accords with [33] who reported that CXCL9 plasma levels were associated with GPT levels and HAI inflammation score, indicating that CXCL9 might have a role in the chronic liver inflammation of HCV-infected patients.

5. Conclusion

In line with the notion of CXCL9 as a key determinant of HCV outcome, our findings confirmed that baseline CXCL9 level significantly correlates not only with SVR, but also with on-treatment viral suppression, which renders it an important prognostic biomarker for viral clearance which may be used to apply patients according to the expectable treatment outcome in HCV genotype 4 patients.

Footnotes

Acknowledgments

Special thanks for Dr. Heba Shawky, Department of therapeutic chemistry, Pharmaceutical Industries and Drug Research Division, and Dr. Reem El shenawy Microbial Biotechnology Department, Genetic Engineering and Biotechnology Research Division National Research Centre, for their help in interpreting the obtained results and in presenting the manuscript in its current form.

References

Cooke

G.S.

Lemoine

Thursz

Gore

Swan

Kamarulzaman

et al., Viral hepatitis and the Global Burden of Disease: A need to regroup, J Viral Hepat 20 (2013), 600–601.

Rehermann

, Hepatitis C virus versus innate and adaptive immune responses: A tale of coevolution and coexistence, J Clin Invest 119 (2009), 1745–1754.

Razavi

ElKhoury

A.C.

Elbasha

Estes

Pasini

Poynard

et al., Chronic hepatitis C virus (HCV) disease burden and cost in the United States, Hepatology 57 (2013), 2164–2170.

El-Serag

H.B.

, Epidemiology of viral hepatitis and hepatocellular carcinoma, Gastroenterology 142 (2012), 1264–1273.

Mohd Hanafiah

Groeger

Flaxman

A.D.

and Wiersma

S.T.

, Global epidemiology of hepatitis C virus infection: New estimates of age-specific antibody to HCV, Hepatology 57 (2013), 1333–1342.

Garrone

Fluckiger

A.C.

Mangeot

P.E.

Gauthier

Dupeyrot-Lacas

Mancip

Cangialosi

Du Chéné

LeGrand

Mangeot

Lavillette

Bellier

Cosset

F.L.

Tangy

Klatzmann

and Dalba

, A prime-boost strategy using virus-like particles pseudotyped for HCV proteins triggers broadly neutralizing antibodies in macaques, Sci Transl Med 3 (2011), 94ra71.

Abdelwahab

S.F.

, Cellular immune response to hepatitis-C-virus in subjects without viremia or seroconversion: Is it important? Infect Agent Cancer 16 (2016), 11–23.

Chin

J.L.

Nicholas

R.M.

Russell

Carr

Connell

Stewart

and McCormick

P.A.

, Spontaneous clearance of hepatitis C infection after liver transplantation from IL28B rs12979860 CC donors, Eur J Gastroenterol Hepatol 24 (2012), 1110–1112.

Logvinoff

Major

M.E.

Oldach

Heyward

Talal

Balfe

Feinstone

S.M.

Alter

Rice

C.M.

and McKeating

J.A.

, Neutralizing antibody response during acute and chronic hepatitis c virus infection, Proc Natl Acad Sci USA 101 (2004), 10149–10154.

10.

Merat

S.J.

Molenkamp

Wagner

Koekkoek

S.M.

van de Berg

Yasuda

Böhne

Claassen

Y.B.

Grady

B.P.

Prins

Bakker

A.Q.

de Jong

M.D.

Spits

Schinkel

and Beaumont

, Hepatitis C virus broadly neutralizing monoclonal antibodies isolated 25 years after spontaneous clearance, PLoS One 11 (2016), e0165047.

11.

Bartolomé

Castillo

Quiroga

J.A.

and Carreño

, Interleukin-28B polymorphisms and interferon gamma inducible protein-10 serum levels in seronegative occult hepatitis C virus infection, J Med Virol 88 (2016), 268–274.

12.

Farci

Alter

H.J.

Wong

D.C.

Miller

R.H.

Govindarajan

Engle

Shapiro

and Purcell

R.H.

, Prevention of hepatitis C virus infection in chimpanzees after antibody-mediated in vitro neutralization, Proc Natl Acad. Sci USA 91 (1994), 7792–7796.

13.

Washburn

M.L.

Bility

M.T.

Zhang

Kovalev

G.L.

Buntzman

Frelinger

J.A.

Barry

Ploss

Rice

C.M.

and Su

, A humanized mouse model to study hepatitis C virus infection, immune response, and liver disease, Gastroenterology 140 (2011), 1334–1344.

14.

Ghany

M.G.

Kleiner

D.E.

Alter

Doo

Khokar

Promrat

et al., Progression fibrosis in chronic hepatitis C, Gastroenterology 124 (2003), 97–104.

15.

Diago

Castellano

Garcia-Samaniego

Perez

Fernandez

Romero

et al., Association of pretreatment serum interferon {gamma} inducible protein 10 levels with sustained virological response to peginterferon plus ribavirin therapy in genotype 1 infected patients with chronic hepatitis C, Gut 55 (2006), 374–379.

16.

Irvine

K.M.

Wockner

L.E.

Hoffmann

et al., Multiplex serum protein analysis identifies novel biomarkers of advanced fibrosis in patients with chronic liver disease with the potential to improve diagnostic accuracy of established biomarkers, PLoS ONE 11 (2016), e0167001.

17.

Jimenez-Sousa

M.A.

Gomez-Moreno

A.Z.

Pineda-Tenor

et al., CXCL9-11 polymorphisms are associated with liver fibrosis in patients with chronic hepatitis C: A cross-sectional study, Clin. Trans. Med. 6 (2017), 26–35.

18.

Ostojic

Markotic

Kelava

and Mrzljak

, Association between CXCL9/10 polymorphisms and acute rejection of liver allograft, Medicine (Baltimore) 98 (2019), e14612.

19.

O’Brien

J.C.

Rainwater

Y.B.

Malviya

et al., Transcriptional and cytokine profiles identify CXCL9 as a biomarker of disease activity in morphea, J. Invest. Dermatol 137 (2017), 1663–1670.

20.

Zhang

Wang

and Li

, NKG2A is a NK cell exhaustion checkpoint for HCV persistence, Nature Commun 10 (2019), 1507.

21.

El-Shabrawi

M.H.

Sherief

L.M.

Yakoot

Kamal

N.M.

Almalky

M.A.

AbdElgawad

M.M.

Mahfouz

A.A.

Helmy

Kamal

E.M.

Attia

and El-Khayat

H.R.

, Effects of dual sofosbuvir/daclatasvir therapy on, chronic hepatitis C infected, survivors of childhood malignancy, World J Clin Cases 7 (2019), 2247–2255.

22.

Saifan

El-Charabaty

Kleiner

and El-Sayegh

, Effect of hepatitis C virus infection on erythropoiesis in patients on hemodialysis, Int J Nephrol Renovasc Dis 6 (2013), 121–124.

23.

Kanda

Kato

Tsubota

Takada

Nishino

Mikami

Miyamura

Maruoka

Nakamoto

Arai

Fujiwara

Imazeki

and Yokosuka

, Platelet count and sustained virological response in hepatitis C treatment, World J Hepatol 5 (2013), 182–188.

24.

Zeremski

Hooker

Shu

M.A.

Winkelstein

Brown

DesJarlais

D.C.

et al., Induction of CXCR3- and CCR5-associated chemokines during acute hepatitis C virus infection, J Hepatol 55 (2011), 545–553.

25.

Butera

Marukian

Iwamaye

A.E.

Hembrador

Chambers

T.J.

Di Bisceglie

A.M.

et al., Plasma chemokine levels correlate with the outcome of antiviral therapy in patients with hepatitis C, Blood 106 (2005), 1175–1182.

26.

Tabll

El Shenawy

Elsharkawy

and Mohamed

F.Z.

, Serum inducible protein-10 chemokine as a biomarker for clearance of HCV with and without treatment in Egyptian patients, Human Antibodies 27 (2019), 265–273.

27.

Johansson

Talloen

Tuefferd

Darling

Fanning

Fried

M.W.

and Aerssens

, Liver Int 36 (2016), 344–352.

28.

Al-Quaiz

M.N.

Madani

T.A.

and Karawi

M.A.

, The natural history of hepatitis C virus infection, Saudi Med J 24 (2003), S67–S70.

29.

Mateescu

R.B.

Rimbaş

Stăniceanu

Zurac

Dragomir

Voiosu

M.R.

Tătaru

Zota

and Bucurică

, Histological and nonhistological criteria in the evaluation of liver involvement in chronic hepatitis C, Rom J Intern Med 44 (2006), 117–130.

30.

Huang

Shiffman

M.L.

Friedman

et al., A 7-gene signature identifies the risk of developing cirrhosis in patients with chronic hepatitis C, Hepatology 46 (2007), 297–306.

31.

Kotoh

Fukushima

Horikawa

et al., Serum albumin is present at higher levels in alcoholic liver cirrhosis as compared to HCV-related cirrhosis, Exp Ther Med 3 (2012), 72–75.

32.

de Souza

M.M.

Vaisberg

V.V.

Abreu

R.M.

Ferreira

A.S.

da Silva Ferreira

Nasser

P.D.

Paschoale

H.S.

Carrilho

F.J.

and Ono

S.K.

, Relationship with abnormal total bilirubin levels in chronic hepatitis C patients Outcomes from a case-control study, Medicine (Baltimore) 96(11) (2017), e6306.

33.

Lee

L.C.

Huang

Y.H.

C.W.

Wang

Y.J.

Huo

T.I.

Lee

K.C.

and Lin

H.C.

, CXCL9 associated with sustained virological response in chronic hepatitis B patients receiving peginterferon Alfa-2a therapy: A pilot study, PLoS One 8(10) (2013), e76798.

CXCL9 chemokine level is associated with spontaneous clearance and sustained virological response in Egyptian Chronic Hepatitis C patients receiving direct acting antivirals

Abstract

BACKGROUND:

AIM:

MATERIALS AND METHODS:

RESULTS:

CONCLUSION:

Keywords

1. Introduction

2. Patients and methods

2.2 Blood sampling

2.3 Biochemical and hematological analysis

2.4 HCV-RNA quantification

2.5 CXCL9 quantification

2.6 Statistical analysis

Table 1 The hematological parameters profile among cohort study

3.1 The hematological profile among studied cohorts

3.3 Levels of CXCL9 in the different groups under the study

3.4 The correlation between CXCL9 and some biochemical parameters

4. Discussion

5. Conclusion

Footnotes

Acknowledgments

References

Table 1
The hematological parameters profile among cohort study