Hamid Reza Rahimi; Bahador Sarkari; Tahereh Mohammadzadeh; Seyed Mahmoud Sadjjadi
Volume 8, Issue 4 , December 2011, , Pages 236-243
Abstract
Background: Cystic echinococcosis (CE), also known as echinococcosis/hydatidosis, is one of the most important parasitic diseases in the world. It enhances both humoral and cellular (Th1 and Th2) responses in infected host. Different antigens of the worm may favor the Th1 or Th2 immune responses in CE ...
Read More
Background: Cystic echinococcosis (CE), also known as echinococcosis/hydatidosis, is one of the most important parasitic diseases in the world. It enhances both humoral and cellular (Th1 and Th2) responses in infected host. Different antigens of the worm may favor the Th1 or Th2 immune responses in CE patients. Objective: To evaluate the humoral and cellular immune responses of Balb/c mice against the crude and excretory/ secretory (E/S) antigens of in vitro reared Echinococcus granulosus adult worms. Methods: A total of 20 Balb/c mice divided into 5 groups of 4 mice each. Three groups of mice (n=4) were immunized with crude, E/S and an immunodominant antigen of in vitro reared Echinococcus granulosus adult worms on day 1 and 28. The fourth and the fifth groups were negative control groups and received PBS plus adjuvant, or nothing, respectively. Two weeks after the second injection, the mice were killed and their blood was collected for determining antibody responses, and their spleens were employed for proliferation assay. Total IgG were measured by indirect ELISA. Spleen cells of immunized mice were cultivated and exposed to different antigens of adult worms including E/S and crude antigens. Level of IFN-γ, IL-12, IL-4 and IL-10 were measured in the recovered cell culture supernatants by capture ELISA. Results: Total IgG assay showed the highest level of antibody produced in mice immunized with crude antigens. Proliferation assay showed a statistically significant production of cytokines in the mice immunized with crude antigens (p<0.05). The highest levels of IFN-γ, IL12 and IL-4 were produced in mice immunized with crude antigen of the in vitro reared Echinococcus granulosus adult worms followed by E/S antigens. Immunodomonant antigen induced the lowest levels of cytokines (IL-12, IFN-γ, IL-4 and IL-10) in immunized mice. Conclusion: A significant levels of Th1 related cytokines (IFN-γ and IL-12) were produced in Balb/c mice immunized with crude antigen of the in vitro reared Echinococcus granulosus adult worms.
Behnam Mohammadi-Ghalehbin; Gholam Reza Hatam; Bahador Sarkari; Mehdi Mohebali; Zabih Zarei; Mansoureh Jaberipour; Shahab Bohlouli
Volume 8, Issue 4 , December 2011, , Pages 244-250
Abstract
Background: Visceral leishmaniasis (VL) is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis ...
Read More
Background: Visceral leishmaniasis (VL) is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis (CVL). Objective: In this survey a Fucose-Mannose Ligand (FML) ELISA, using native L. infantum antigen, was developed and its validity for detection of infected dogs in comparison with direct agglutination test (DAT) and PCR was evaluated. Methods: Blood samples of sixty ownership dogs (≤ 3 years old) were collected from Meshkin-shahr district in Ardabil province, North-west of Iran. Sera were separated for serological assays (DAT and FMLELISA) and the buffy coats were collected for molecular evaluation. Results: Two out of the 60 (3.33%) samples were found to be positive (antibody titer of ≥ 1/320) in DAT while seven of the 60 (11.66%) samples were positive by FML-ELISA. Nine out of 60 (15%) buffy coat samples showed a band about 680 bp indicative of L. infantum in PCR. Three out of 60 dogs had Kala-azar symptoms and were positive by PCR and FML-ELISA, while two of these three dogs had antibody titers >1/320 in their serum samples. The sensitivity and specificity of FML-ELISA for the detection of CVL in both symptomatic and asymptomatic dogs were found to be 77.8% and 100%, respectively. Conclusion: Considering the acceptable sensitivity and high specificity of FMLELISA, use of this serological method can be recommended for epidemiological surveys of CVL.