Institute of Massachusetts General Hospital, Massachusetts Institute of
Technology and Harvard University, 400 Technology Square, Cambridge
2 Department of Immunology and Biotechnology, Tropical Medicine Research Institute. National Centre for Research, Khartoum, Sudan
3 National Centre for Research, Ministry of Higher Education and Scientific Research, Khartoum, Sudan
Received: March 26, 2017
Accepted: June 8, 2017
Mediterr J Hematol Infect Dis 2017, 9(1): e2017042 DOI 10.4084/MJHID.2017.042
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(Hp) is an acute phase protein that binds the free hemoglobin (Hb),
thus preventing iron loss and renal damage. Hp also has antioxidative
and immunomodulatory properties. Three Hp phenotypes have been
identified in human: Hp1-1, Hp2-1, and Hp2-2. Hp polymorphisms have
been related to susceptibility of various diseases. In this study, we
aimed to assess the possible association of Hp phenotypes polymorphism
to Schistosoma parasites infection in central Sudan. We have investigated the Hp phenotypes polymorphism distribution in the serum of 125 (93 S. mansoni, 13 S. haematobium
and 19 infected with both ‘’co-infection’’) parasitologically confirmed
infected individuals and 208 healthy individuals served as control. Hp
phenotypes have been determined by polyacrylamide gel electrophoresis
followed by benzidine staining. Our study revealed that Hp1-1
percentage frequency was significantly higher in infected individuals
than healthy control individuals 51% and 26% respectively. Our data
suggest that Hp1-1 phenotype may upsurge the susceptibility to Schistosoma parasites infection in central Sudan.
Hp phenotypes revealed different anti-inflammatory, immunomodulatory and anti-oxidative properties that have clinical consequences in different pathologies including cancer, infections and also the lifespan expectancy.[6-9] As an immunomodulatory, Hp1-1 and Hp2-1 have been reported to affect the T-lymphocyte functions by direct binding to the resting and activated CD4+ and CD8+ T lymphocytes resulting in a strong suppression of induced T-cell proliferation. Furthermore, Hp1-1 and Hp2-1 display strong in vitro inhibitory effect on Th2 cytokine release and subsequently promote Th1 activation over Th2 activation in vivo. Haptoglobin also acts as a potent antioxidant, oxidative damage to DNA induced by hydrogen peroxide have been investigated among the three Hp phenotypes and finding revealed that Hp1-1 has least DNA damage compared to Hp2-1 and Hp2-2.
Schistosomiasis or bilharzia is an infection caused by trematodes (blood flukes) and is widespread in Sudan, especially in the major irrigation systems of the Gezira agriculture scheme between the Blue and White Nile Rivers. Infection in Sudan mainly belongs to two species of the genus Schistosoma: S. mansoni which cause intestinal schistosomiasis and S. haematobium that causes urinary schistosomiasis.[13,14] The proportion of the population infected with schistosomiasis is growing in the endemic areas. The disease has very serious socioeconomic consequence e.g. decreasing work capacity, restricting marriage and occupational mobility. Some studies in Sudan revealed the association of Hp phenotypes polymorphism with infectious and non-infectious diseases.[16-18] Therefore, in this study, we investigated the possible association of Hp phenotypes and susceptibility to Schistosoma parasites infection acquisition in central Sudan. Our finding suggests that the individuals with Hp1-1 are at higher risk of attaining the Schistosoma infection compared to individuals with other Hp phenotypes.
Materials and Methods
Identification of Haptoglobin (Hp) phenotypes. Haptoglobin (Hp) phenotypes were separated in discontinuous polyacrylamide gel electrophoresis (non-reducing) according to Davis and Orenstein method and modified by Linke and was applied using the Mini-V 8.10 system (BRL, Life Technologies Inc, Gaithersburg, USA). In brief, 10 μl of serum was mixed with 4 μl of erythrocyte hemolysate which contains free hemoglobin (Hb) and 5 μl loading buffer, then 10 μl from each prepared mixture was added to each well of 4.7% polyacrylamide gel. After completion of the run, the gel was stained for 10-15 min with benzidine stain.
Statistical analysis. Statistical significance was assessed by Chi-square test to determine the association of Hp phenotypes distribution among infected individuals and healthy control individuals. Statistical analysis and charts preparation were performed using Graphpad Prism version 5.0 (GraphPad Software Inc.).
Results and Discussion
|Figure 2. Overall distribution percentage of different Hp phenotypes among individuals infected with Schistosoma parasites and healthy control individuals. Value of p<0.05 considered significant.|
|Figure 3. Overall distribution percentage of different Hp phenotypes among individuals infected either with S. mansoni, S. haematobium or both “Co-infection” compared to healthy control individuals. Value of p<0.05 considered significant.|
It has been demonstrated that Hp 1-1 possess differentially and extremely greater antioxidant activity than Hp 2-1 and 2-2 which may reflect in clinical implications in various diseases. Indeed Hp1-1 have been associated with malaria infection in some African countries such as Ghana, Cameroon, and Sudan as well.[18,28,30] Some early studies in Sudan also reported that Hp1-1 phenotype is associated with liver disorders including hepatitis B virus (HBV).[16,17] Interestingly a significant increase of Hp concentration following the Schistosoma mansoni cercariae infection in both mice and baboons have been observed and linked to schistosomiasis infection and pathogenesis.[31-33] Our current study findings attest the involvement of Hp in schistosoma infection in particular Hp1-1. Intriguingly Cook and his colleagues found that Schistosoma adult worm expresses antioxidant enzymes to neutralize the effects of reactive oxygen and nitrogen species as a mechanism in evading immune killing. Moreover, Hp1-1 and Hp2-1 playing a critical role in modulating the Th1/Th2 balance and promoting a dominant Th1 cellular response over Th2.[10,11] Th1/Th2 balancing is crucial to fight and combat schistosoma infection by the host as after the penetration of cercariae, and before the onset of egg deposition the immune response is primarily of Th1 type (promoting cellular immunity) and directed against worm antigens. At the onset of egg production, the immune response switches to Th2 type, promoting an antibody response directed preferentially against highly immunogenic egg glycans.
Collectively we suggest that the potent antioxidative activity of the Hp1-1 phenotype may neutralize the effects of reactive oxygen and nitrogen species and therefore support the early stage of Schistosoma parasite in evading immune killing and eventually facilitated the establishment of Schistosoma infection compared to individuals with Hp2-1 or Hp2-2 phenotype. Suppression of Th2 response by Hp1-1 could contribute as well to infection and disease progression.
AcknowledgementsWe would like to thank all participants for agreed to be involved in this study. Our gratitude extended to the Department of Immunology and Biotechnology members of TMRI. This work was supported by the National Centre for Research (NCR), Ministry of Higher Education and Scientific Research- Sudan.
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