Schistosomiasis is a chronic parasitic disease in tropical and is associated with a variety of clinical syndromes that may lead to severe morbidity (Bahgat et al., 2010). Due to control program over the last decade, a decline in the prevalence of human schistosomiasis in Egypt has been reported (Barakat et al., 1998; Bahgat et al., 2010); however, the disease is still endemic in many foci (El-Sahn et al., 2002; Bahgat et al., 2010).
The common used method in the diagnostic of schistosomiasis is stool examination, using the Kato-Katz method (Carvalho et al., 2011); However, this method presents limitation for the detection of positive individuals, when the intensity of infection is low; it is less effective in determining the prevalence in low endemic areas (Feldmeier and Poggensee 1993; Kongs et al., 2001; Carvalho et al., 2011). And a relatively time consuming nature of this method in application for epidemiological assessment and clinical use (Van Etten et al., 1994; Hamilton et al., 1998; Corachan , 2002; Van Dam et al., 2004). And it is difficult to detect ova in rectal biopsy specimens in chronic infections due to the intense fibrosis present around the eggs (Attallah et al., 1999).
Several schistosome serodiagnostic assays designed for the detection of specific anti-schistosome (antibodies) (Doenhoff et al., 1989; Maddison, 1987; Attallah et al., 1999). Prepatent and early infections may not have stimulated a detectable antibody response and Positive tests do not necessarily denote an active (living) infection. Antibody tests are also prone to cross reactions with other infections (Strrouk, 2001).
HCV is a serious public health problem affecting 170 million carriers worldwide. It is a leading cause of chronic hepatitis (Thong et al., 2014). In Egypt, the older generations have a higher HCV prevalence than younger ones. Geographically, areas near the Nile River continue to exhibit very high rates of infection (Sievert et al., 2011). Chronic HCV infection may cause liver cirrhosis and (HCC) over the course of two or more decades (lee et al., 2008), and is the primary cause for liver transplantation worldwide (Thong et al., 2014), There is no vaccine available for prevention of HCV infection due to high degree of strain variation (Ashfaq et al., 2011). A previous study by Albeldawi et al (2010) reported that the patients with risk factors for HCV infection and abnormal liver enzyme levels, HCV infection is probable but not certain.
In the present study 275 serum samples were tested for anti-HCV antibodies and 205/275 only contain anti-HCV antibodies with percent (74.5%) and the remaining 70 samples (25.5 %) were negative for anti-HCV antibodies. Anti-HCV antibodies alone cannot discriminate patients who are infectious from those who have resolved the infection. The active infection confirmed by detected HCV-NS4 antigen in serum samples using ELISA according to Attallah et al., (2012) in which sensitivity and specificity of this method to detect HCV-NS4 antigen were high (90 and 96%, respectively), HCV-NS4 antigen was detected in 205( positive for anti-HCV antibodies) against 32 healthy (negative control) by using ELISA with detection rate 85.0% for CHC patient positive for anti-HCV antibodies, and 0.0% detection rate for healthy control, this mean that 174/205 only has Anti-HCV and HCV-NS4 (HCV patient) , there was extremely statistically significant between (p<0.0001) HCV-NS4 in CHC patient and control . in the previous study by Ghany et al (2009) reported that ALT value differs by age, race, and gender, and by body mass index, it has been suggested that the upper limit of normal (ULN) for ALT should, in fact, be 30 IU/L for men and 19 IU/L for women, but many laboratories continue to set the ULN of ALT at about 40 IU/L. In the present study, 174 HCV have high increased in transaminase enzyme as ALT, AST, and there was extremely statistically significant between (p<0.0001) transaminase enzyme (ALT, AST) in CHC patient and negative control.
As is customary the transaminase enzymes are biomarker of liver health. Elevated serum levels of ALT during chronic hepatitis C are associated with an increased risk of liver fibrosis progression (Hui et al., 2003; Maasoumy and Wedemeyer, 2012); lower progression rates of fibrosis were reported in patients with normal ALT levels (Mathurin et al., 1998; Maasoumy and Wedemeyer, 2012); However, there are reports of marked fibrosis (5%-30%) and even cirrhosis (1.3%) in persons with normal ALT values (Ghany et al., 2009).
Normal ranges of ALP were 40-129 U/L for males and 35-104 U/L for females, ALP is elevated in a variety of diseases as bone disease, bile duct obstructed and liver disease (Bodlaj et al., 2010), and in current study ALP was elevated in CHC patient and there was statistically significant between (p<0.01) ALP in CHC patient and negative control. In the present study, it showed a highly decrease in the values of serum albumin and the albumin protein is produced in the liver but there was extremely statistically significant between (p<0.0001) serum albumin in CHC patients and negative control and it showed a highly decrease in the platelets for the CHC patients compare to healthy individuals (control group) but there was no statistically significant between (p>0.05) platelets in CHC patients and negative control, Furthermore, T. Bilirubin was elevated in CHC patient and there was statistically significant between (p<0.05) T. Bilirubin in CHC patient and control. There was no statistically significant (p>0.05) between INR-prothrombin time in CHC patient and control.
A Previous study by Attallah et al (1999) reported that Schistosoma circulating antigens were used for the detection of active infection. And the detection of Schistosoma antigens was initially based on the use crude soluble egg antigens (SEA) and soluble adult worm’s protein (SWAP) (Dunne et al., 1984). And another study reported that the circulating antigens could be released from the schistosome surface or gut to the blood circulation of infected host and consequently excreted in urine (Deelder et al., 1994; Attallah et al., 1998). Several investigators have isolated and characterized many of the schistosomiasis antigens in different developmental stages of the parasite that have a potential application in immuno diagnosis (Attallah et al., 1999). And more than 100 schistosome antigens have been identified (Siddiqui et al., 2011).
In The present study we aimed to detect the 63-KDa circulating antigen of Schistosoma parasites in serum samples by SDS-PAGE, immunoblotting, and ELISA technique, The antigenic target was identified by western blotting technique by using specific Mab; an intense sharp band was appeared in serum samples of infected patients with S. mansoni at 63-kDa, and no reaction with healthy sample. Consistent with these findings previously study by Attallah et al (1999) also identified a 63-kDa antigen in different extracts of Schistosoma mansoni cercariae, adult worms, and eggs and in the urine of S.mansoni-infected individuals by specific monoclonal antibody in urine patient sample.
ELISA is a serological test useful for epidemiological studies, due to its high sensitivity for the diagnosis, depending on the S.mansoni antigen used in the test (Sorgho et al., 2002; Ishida et al., 2003; Alarcón de Noya et al., 2007;Luo et al., 2009; Carvalho et al.,2011), and the possibility for automating the process (Carvalho et al.,2011), previous study Attallah et al (1999)used the Fast Dot-ELISA as a diagnostic tool for the detection of 63-kD circulating antigen and sensitivity of this assay was 92% among proven S. mansoni-infected individual, the specificity was 84%, PPV = 94%, and NPV = 81%.
In the current study, the cutoff of 63-KDa of Schistosoma antigen was calculated, and the cutoff level = 0.33, mouse monoclonal antibody specific to S. mansoni was used as a probe in ELISA to detect 63-kDa S. mansoni antigen in serum samples from CHC patients, the 63-kDa S. mansoni antigen was detected in 118 out of total 174 chronic hepatitis C patients with detection rate (67.8%) All of the 32 controls were negative for 63-kDa S. mansoni antigen with detection rate (0.0 %) and there was extremely statistically significant between (P<0.0001) 63-KDa S.mansoni antigen in CHC patient and control.
In the present study we determined the relation between 63-KDa S. mansoni antigen and laboratory parameters in the 118 serum samples from CHC patients determined, and serum ALT levels tended to increase as the level of 63-KDa antigen increased and there was extremely statistically significant between 63-KDa and ALT, serum AST levels tended to increase as the level of 63-KDa antigen increased and there was extremely statistically significant between 63- KDa and AST.
Furthermore, there was negative correlation between serum albumin and 63-KDa antigen, but there was extremely statistically significant between 63-KDa and serum albumin, serum ALP levels tended to increase as the level of 63-KDa antigen increased but this was not statistically significant, total bilirubin levels tended to increase as the level of 63-KDa antigen increased but this was not statistically significant, there was negative correlation between PLTs count and 63-KDa antigen and this was not statistically significant, INR-prothrombin time levels tended to increase as the level of 63-KDa antigen increased but this was not statistically significant.
Both HCV and schistosomiasis are highly endemic in Egypt and coinfection is frequently encountered (Abdel-Rahman et al., 2013). The prevalence of HCV/S.mansoni association ranged from 0.8 to 50.0% among the studies, with the highest ranges in Egypt (10 to 50%) (Van- Lume et al., 2013). A previous study by Blenton et al (2002) mentioned that Coinfections might influence disease expression. HCV has been implicated in some studies as a factor influencing the severity of schistosomiasis (Mohamed et al., 1998; Blenton et al., 2002). And, in turn, an influence of schistosomiasis on HCV severity has been suggested (Kamal et al., 2000; (Blenton et al., 2002). Another study suggests that Schistosomiasis weaken anti- HCV immune responses and worsen liver disease (Kamal et al., 2000; Kamal et al., 2001; Osada and Kanazawa, 2011).
It is worthy to notice that another study by Abdel-Rahman et al (2013) reported that HCV/schistosomiasis coinfected patients have more rapid progression of hepatic fibrosis than those with HCV mono-infection encountered, the effect of such coinfection on hepatic and response to therapy remain unclear.
In light of the previous by Bahgat et al (2010) real time PCR findings from, soluble egg antigen (SEA) should be considered as a potential stimulatory factor for HCV RNA that may have influenced the early detection of HCV RNA as SEA can stimulate viral replication. The higher morbidity that is observed in patients coinfected with schistosomiasis and HCV is related, at least in part, to direct stimulation of viral replication by SEA. Another study by Van–lume et al (2013) reported that when schistosomiasis and hepatitis C association is established, the clinical course develops into severe hepatocellular damage. Viral persistence and hepatic cirrhosis can develop faster than in mono-infected people.
The association between schistosomiasis and hepatitis C has been studied by many investigators due to its important but, the object of current study was undertaken to determine a correlation between HCV and schistosomiasis infection in relation to evaluation of HCV-NS4 in CHC patient only and S.mansoni/HCV co-infection , from our finding r = 0.407, this means that, there was positive association was observed between 63 KD-a S. mansoni antigen and HCV-NS4 antigen, and there was extremely statistically significant between (p < 0.0001) 63 KD-a S. mansoni antigen and HCV-NS4 antigen, it is of interest that our study is the first to focus on the relation between 63-KDa S. mansoni antigen and HCV-NS4 antigen.
In present study, we made comparison between investigated blood markers in CHC patient co-infected and HCV only, from our finding there was increase in activities of ALT, AST in CHC patients co-infected than HCV only, On the other hand, there was decrease in serum albumin level, platelets count for CHC patients co-infected compared with non-infected with S. mansoni group and there was extremely statistically significant between (p<0.05) AST in CHC patients (S.mansoni/HCV) co-infected and non-infected with S. mansoni (HCV only), but this was not statistically significant between ALT in CHC patients (S.mansoni/HCV) co-infected and non-infected with S. mansoni (HCV only).
The transaminase enzymes are indicator of liver fibrosis, this may be that co-infection accelerate tissue damage and liver fibrosis, there was no statistically significant between ALP in CHC patients (S.mansoni/HCV) co-infected and non-infected with S. mansoni (HCV only), this was no statistically significant between bilirubin in CHC patients (S.mansoni/HCV) co-infected and non-infected with S. mansoni (HCV only), there was no statistically significant between Albumin in CHC patients (S.mansoni/HCV) co-infected and non-infected with S. mansoni (HCV only), there was no statistically significant between PLTs count in CHC patients (S.mansoni/HCV) co-infected and non-infected with S. mansoni (HCV only). In the previous study by reported that S.mansoniinfection is known to induce hepatocellular injury, which in turn, leads to the release of enzymes from the injured hepatic cells into the blood circulation (Dkhil, 2014). In current study, showed that an increase in the level of HCV-NS4 antigen in patient with S.mansoni/HCV than level of HCV-NS4 antigen in patient with HCV only.
In conclusion, the 63-KDa was the antigenic component of S.mansoni, our study showed increased activity in transaminase enzyme and decreased activity in Albumin and PLTs in S.mansoni/HCV coinfection than HCV only, and 63-KDa antigen S.mansoni has a positive correlation with HCV-NS4 antigen