Giardia duodenalis assemblages in Egyptian children with diarrhea.

Giardia duodenalis is considered the most common cause of parasitic diarrhea worldwide. Genetic studies revealed that at least eight assemblages (A-H) exist. Of these assemblages, A and B are found primarily in human beings and occasionally in animals. The association between clinical symptoms and G. duodenalis assemblages is controversial. The aim of the present study was to determine the assemblages of G. duodenalis prevalent among Egyptian children with diarrhea. Therefore, 96 positive stool samples for Giardia by light microscopy were subjected to multilocus genotyping targeting the triose phosphate isomerase (tpi), ß-giardin (bg), and glutamate dehydrogenase (gdh) genes. Amplified polymerase chain reaction (PCR) products were then purified, sequenced, and aligned with reference strains to determine the assemblages of the Giardia isolates. Out of the 96 microscopically positive stool samples for Giardia, 77 (80 %) were successfully amplified and sequenced at least at one locus. Of these, 21 (27.3 %) were shown to be assemblage A, 54 (70.1 %) assemblage B, while discordant sequence typing results were observed in 2 (2.6 %) samples. AII was the predominant subassemblage of assemblage A, while it was generally difficult to further classify assemblage B. It was concluded that infection with assemblage B was more common than that with assemblage A. No associations between epidemiological information and assemblage were detected, except with age. Although infections with assemblage B were more frequently associated with abdominal pain and acute diarrhea than with assemblage A, the difference was not statistically significant.

Use of multiplex real-time PCR for detection of common diarrhea causing protozoan parasites in Egypt.

Diarrhea is an important cause of morbidity and mortality, worldwide. Giardia intestinalis, Cryptosporidium spp., and Entamoeba histolytica are the most common diarrhea-causing parasitic protozoa. Diagnosis of these parasites is usually performed by microscopy. However, microscopy lacks sensitivity and specificity. Replacing microscopy with more sensitive and specific nucleic acid based methods is hampered by the higher costs, in particular in developing countries. Multiplexing the detection of more than one parasite in a single test by real-time polymerase chain reaction (PCR) has been found to be very effective and would decrease the cost of the test. In the present study, stool samples collected from 396 Egyptian patients complaining of diarrhea along with 202 faecal samples from healthy controls were examined microscopically by direct smear method and after concentration using formol-ethyl acetate. Frozen portions of the same samples were tested by multiplex real-time for simultaneous detection of E. histolytica, G. intestinalis, and Cryptosporidium spp. The results indicate that among diarrheal patients in Egypt G. intestinalis is the most common protozoan parasite, with prevalence rates of 30.5 and 37.1 %, depending on the method used (microscopy vs. multiplex real-time PCR). Cryptosporidium spp. was detected in 1 % of the diarrheal patients by microscopy and in 3 % by real-time PCR. While E. histolytica/dispar was detected in 10.8 % by microscopy, less than one fifth of them (2 %) were found true positive for Entamoeba dispar by real-time PCR. E. histolytica DNA was not detected in any of the diarrheal patients. In comparison with multiplex real-time PCR, microscopy exhibited many false positive and negative cases with the three parasites giving sensitivities and specificities of 100 and 91 % for E. histolytica/dispar, 57.8 and 85.5 % for G. intestinalis, and 33.3 and 100 % for Cryptosporidium spp.

Pegylated interferon alpha-2b plus ribavirin in patients with genotype 4 chronic hepatitis C: The role of rapid and early virologic response.

UNLABELLED: In patients chronically infected with hepatitis C virus (HCV) genotype 4, the optimum duration of therapy and the predictors of sustained virologic response (SVR) have not been adequately determined. In this study, 358 patients with chronic hepatitis C genotype 4 were randomly assigned to pegylated interferon (PEG-IFN) alpha-2b (1.5 mug/kg/week) plus oral ribavirin (10.6 mg/kg/day) for a fixed duration of 48 weeks (control group, n = 50) or for a variable duration (n = 318). In the variable-duration group, patients with undetectable HCV RNA at week 4 were treated for 24 weeks (group A, n = 69), patients with undetectable HCV RNA at week 12 were treated for 36 weeks (group B, n = 79), and the rest of the patients were treated for 48 weeks (group C, n = 160). The primary endpoint was SVR (undetectable HCV RNA 24 weeks after treatment cessation). Groups A-C and the control group had SVR rates of 86%, 76%, 56%, and 58%, respectively. After the study was controlled for predictors, a low baseline histologic grade and stage were associated with SVR (P < 0.029) in all groups. In addition, among patients in group C, older age (P = 0.04), a higher baseline body mass index (P = 0.013), and low baseline HCV RNA (P < 0.001) were also associated with SVR attainment. The incidence of adverse events and the rate of discontinuation were higher in patients in the variable-duration and fixed-duration groups treated for 48 weeks. CONCLUSION: In patients with chronic hepatitis C genotype 4 and undetectable HCV RNA at weeks 4 and 12, treatment with PEG-IFN alpha-2b and ribavirin for 24 weeks and 36 weeks, respectively, is sufficient.

Immune modulation by helminthic infections: worms and viral infections.

Helminthic infections occur worldwide, especially in developing countries. About one-quarter of the world's population, 1.5 billion, are infected with one or more of the major soil-transmitted helminths, including hookworms, ascarids, and whipworms. Schistosomes infect more than 200 million people worldwide with 600 million at risk in 74 countries. The interaction between helminths and the host's immune system provokes particular immunomodulatory and immunoregulatory mechanisms that ensure their survival in the host for years. However, these changes might impair the immunological response to bystander bacterial, viral, and protozoal pathogens and to vaccination. Modulation of the immune system by infection with helminthic parasites is proposed to reduce the levels of allergic responses and to protect against inflammatory bowel disease. In this review, we summarize the immunological milieu associated with helminthic infections and its impact on viral infections, mainly hepatitis B virus, hepatitis C virus, and human immunodeficiency virus in humans and experimental animals.

Specific cellular immune response and cytokine patterns in patients coinfected with hepatitis C virus and Schistosoma mansoni.

Patients coinfected with hepatitis C virus (HCV) and Schistosoma mansoni show high incidence of viral persistence and accelerated fibrosis. To determine whether immunological mechanisms are responsible for this alteration in the natural history of HCV, the HCV-specific peripheral CD4(+) T cell responses and cytokines were analyzed in patients with chronic hepatitis C monoinfection, S. mansoni monoinfection, or HCV and S. mansoni coinfection. An HCV-specific CD4(+) proliferative response to at least 1 HCV antigen was detected in 73.3% of patients infected with HCV, compared with 8.6% of patients coinfected with HCV and S. mansoni. Stimulation with HCV antigens produced a type 1 cytokine profile in patients infected with HCV alone, compared with a type 2 predominance in patients coinfected with HCV and S. mansoni. In contrast, there was no difference in response to schistosomal antigens in patients infected with S. mansoni alone, compared with those coinfected with HCV and S. mansoni. These findings suggest that the inability to generate an HCV-specific CD4(+)/Th1 T cell response plays a role in the persistence and severity of HCV infection in patients with S. mansoni coinfection.

Acute hepatitis C without and with schistosomiasis: correlation with hepatitis C-specific CD4(+) T-cell and cytokine response.

BACKGROUND & AIMS: Immune responses during the first few months of acute hepatitis C virus (HCV) infection seem crucial for viral control, but the relationship of these responses to natural history is poorly characterized. METHODS: This prospective study investigated the HCV-specific CD4(+) and cytokine responses in patients with acute HCV hepatitis with or without Schistosoma mansoni coinfection, a parasitic infection with T helper (Th) 2 immune bias. HCV-specific CD4(+) proliferative responses and cytokine production in peripheral blood mononuclear cells were correlated with liver biopsy results at 6 months and at the end of follow-up. RESULTS: Whereas 5 of 15 patients with HCV alone recovered from acute HCV, all (17 of 17) patients with S. mansoni coinfection progressed to histologically proven chronic hepatitis. Coinfected patients had either absent or transient weak HCV-specific CD4(+) responses with Th0/Th2 cytokine production. The magnitude of the HCV-specific CD4(+) response at week 12 was inversely correlated with the fibrosis progression rate in chronically infected patients. CONCLUSIONS: Patients with acute hepatitis C and schistosomiasis coinfection cannot clear viremia and show rapid progression once chronic infection is established. This rapid progression is associated with a strong Th2 response in peripheral immune responses, suggesting that early development of vigorous Th1 responses not only facilitates clearance but delays disease progression.