Increased risk for Entamoeba histolytica infection and invasive amebiasis in HIV seropositive men who have sex with men in Taiwan

Background

Incidence of Entamoeba histolytica infection and clinical manifestations and treatment response of invasive amebiasis (IA) in HIV-infected patients have rarely been investigated before.

Methodology/Principal Findings

At the National Taiwan University Hospital, medical records of HIV-infected patients who received a diagnosis of IA between 1994 and 2005 were reviewed. The incidence of amebiasis was investigated in serial blood and stool samples from 670 and 264 HIV-infected patients, respectively, using serological and specific amebic antigen assays. DNA extracted from stool samples containing E. histolytica were analyzed by PCR, sequenced, and compared. Sixty-four (5.8%) of 1,109 HIV-infected patients had 67 episodes of IA, and 89.1% of them were men having sex with men (MSM). The CD4 count at diagnosis of IA was significantly higher than that of the whole cohort (215 cells/µL vs. 96 cells/µL). Forty episodes (59.7%) were liver abscesses, 52 (77.6%) colitis, and 25 (37.3%) both liver abscesses and colitis. Fever resolved after 3.5 days of metronidazole therapy (range, 1–11 days). None of the patients died. The incidence of E. histolytica infection in MSM was higher than that in other risk groups assessed by serological assays (1.99 per 100 person-years [PY] vs. 0 per 100 PY; p<0.0001) and amebic antigen assays (3.16 per 100 PY vs. 0.68 per 100 PY; p = 0.12). In multiple logistic regression analysis, only MSM was significantly associated with acquisition of E. histolytica infection (adjusted odds ratio, 14.809; p = 0.01). Clustering of E. histolytica isolates by sequencing analyses from geographically-unrelated patients suggested person-to-person transmission.

Conclusions/Significance

HIV-infected MSM were at significantly higher risk of amebiasis than patients from other risk groups. Despite immunosuppression, amebic liver abscesses and colitis responded favorably to treatment.     

Pathophysiology of thrombocytopenia associated with HIV infection in homosexual men. A preliminary report

The pathophysiology of HIV associated immune thrombocytopenia (HIV-ITP) and its response to AZT was investigated. Using autologous 111Indium-labelled platelets, platelet kinetic were analysed in two patient groups. Group 1 (untreated) was comprised of 13 patients with HIV-ITP. Group 2 (AZT) was comprised of 6 patients with a history of HIV-ITP prior to starting AZT. These patients were studied following a rise in their platelet count on AZT. Platelet survivals in both groups were shortened compared to controls, however there was no significant difference between the 2 groups. However platelet turnover rates were significantly depressed in Group 1 compared to Group 2 (P less than 0.05) and control values (P less than 0.05). The platelet count correlated with platelet turnover in Group 1 but not in AZT treated patients. Thus patients with HIV-ITP appear to have both shortened survivals and suppressed platelet production. The mechanism of the latter is unknown, but the increased turnover rate seen with AZT suggests it is a direct or indirect effect of the HIV virus.     

Entamoeba histolytica and/or Entamoeba dispar: infection frequency in HIV+/AIDS patients in Mexico city

The objective of this work was to evaluate the frequency of Entamoeba histolytica/Entamoeba dispar intestinal infection in HIV+/AIDS subjects and their HIV- close relatives or sexual partners. Enteric parasites were investigated in stool samples by microscopic examination and E. histolytica and E. dispar were identified by PCR. We found by microscopic analysis in HIV+/AIDS group that the E. histolytica/E. dispar complex was present in 5.9% of the members, while in the HIV- group was 2.9%. With PCR we found that the E. histolytica prevalence was 25.3% in the HIV+/AIDS group and 18.5% in the HIV-group. The difference in the results obtained with the microscopic and PCR is due to the different sensibility of the procedures. Besides, we found patients who were infected with E. histolytica in both groups were asymptomatic cyst passers. Our results suggest that E. histolytica strains prevalent in the studied community appear to be of low pathogenic potential.     

Introns of Entamoeba histolytica and Entamoeba dispar.

The genome of Entamoeba histolytica is considered to possess very few intervening sequences (introns), as only 5 intron-containing genes from this protozoan parasite have been reported so far. However, while sequencing a number of genomic contigs as well as three independent genes coding for ribosomal protein L27a, we have identified 9 additional intron-containing genes of E. histolytica and the closely related species Entamoeba dispar, indicating that introns are more common in these organisms than previously suggested. The various amoeba introns are relatively short comprising between 46 and 115 nucleotides only and have a higher AT-content compared to the corresponding exon sequences. In contrast to higher eukaryotes, amoeba introns do not contain a well-conserved branch point consensus, and have extended donor and acceptor splice sites of the sequences G     

Experimental infection of rhesus monkeys with Entamoeba histolytica mimics human infection

Experimental infection of Entamoeba histolytica was successfully established in rhesus monkeys (Macaca mulatta). Parasites of proven pathogenicity maintained through liver passage in hamsters were used in this study. Laparotomized monkeys were inoculated intracecally/intrahepatically with trophozoites, or by oral administration of cysts (2000 cysts/ml). The cysts were isolated from an infected human stool sample. Formation of typical amebic lesions in the inoculated tissues along with alterations in hematologic indices were studied in the infected monkeys. All the experimentally inoculated monkeys showed leukocytosis and mild neutrophilia, while the hemoglobin content and levels of blood sugar and blood urea remained unaltered during the post-infection period. Specific antiamebic antibodies were readily detectable in post-infected sera.     

Changes in sexual behaviour and the fall in incidence of HIV infection among homosexual men.

To investigate the epidemiology and normal course of infection with HIV the prevalence and incidence of the infection were studied among two cohorts of homosexual men in Amsterdam in 1980-7. The cumulative incidence of infection increased from a weighted 2.2% in 1980 to 39.0% in 1987. The estimated yearly incidence of HIV was 3.0% in 1981, rose to 8.8% in 1984, and fell gradually to 0% in 1987. During the study the sexual behaviour of the cohorts was examined. The number of men with whom anopenetrative intercourse was practised fell from a mean of 10.6 to 1.4 for those positive for HIV antibody, whereas the number with whom anoreceptive intercourse was practised fell from a mean of 3.7 to 0.5 for those negative for the antibody. In addition, there was a reduction in the number of cases of hepatitis B and syphilis among men in general. The decline in infection with HIV was assumed to be linked to changes in sexual behaviour. Such changes practised early in the course of the epidemic probably had a strong effect on the number of cases of AIDS among homosexual men in Amsterdam.     

Entamoeba histolytica and Entamoeba dispar: prevalence infection in a rural Mexican community

The frequency of Entamoeba histolytica and Entamoeba dispar infection was analyzed in a rural community in the state of Morelos, Mexico, through PCR technique by using specie specific primer. The E. histolytica specie was detected in 33 of 290 analyzed stool samples (11.4%), E. dispar specie was observed in 21 samples (7.2%) and both species of Entamoeba were detected in seven samples (2.4%). So a higher E. histolytica than E. dispar frequency infection was detected (13.8 versus 9.6%). Even though in our design we did not considered the follow-up of included individuals, the absence of invasive amebiasis cases in the studied population during our stay in town was unexpected.     

Molecular biology research to benefit patients with Entamoeba histolytica infection

The development of molecular microbiology has made it possible for us to deepen our understanding of the pathogenesis of amebiasis. Research using the trophozoite form of Entamoeba histolytica has clearly shown us the importance of the interface between the parasite and host cells in vitro. Immuno‐pathogenesis after excystation was similarly well advanced by the use of a novel murine model of amebic colitis. However, it is still challenging to apply these findings to clinical and epidemiological settings. This is mainly because of the lack of a complete infection animal model of amebiasis by oral–fecal infection. Moreover, in vitro experiments have predominantly been performed using the same axenic cultured strain HM‐1: IMSS isolated about 50 years ago, whereas highly diverse strains are prevalent all over the world. Translational research informed by clinical observations has the greatest potential for the development of effective interventions. Here, we highlight discoveries of the experiments designed from cohort observation and discuss remaining problems to be solved.     

Codon usage in Entamoeba histolytica.

The codon usage of 10 E. histolytica genes comprising 4455 codons was analysed. The codon usage revealed an extremely biased use of synonymous codons with a preference for NNU (44%) and NNA (41.4%) codons. Codons CGG (arg), AGG (arg) and CCG (pro) were absent in the E. histolytica genes examined. The codon usage of E. histolytica resembled that of Plasmodium falciparum.     

Pyruvate-to-ethanol pathway in Entamoeba histolytica.

The pyruvate-to-ethanol pathway in Entamoeba histolytica is unusual when compared with most investigated organisms. Pyruvate decarboxylase (EC 4.1.1.1), a key enzyme for ethanol production, is not found. Pyruvate is converted into acetyl-CoA and CO2 by the enzyme pyruvate synthase (EC 1.2.7.1), which has been demonstrated previously in this parasitic amoeba. Acetyl-CoA is reduced to acetaldehyde and CoA by the enzyme aldehyde dehydrogenase (acylating) (EC 1.2.1.10) at an enzyme activity of 9 units per g of fresh cells with NADH as a reductant. Acetaldehyde is further reduced by either a previously identified NADP+-linked alcohol dehydrogenase or by a newly found NAD+-linked alcohol dehydrogenase at an enzyme activity of 136 units per g of fresh cells. Ethanol is identified as the product of soluble enzymes of amoeba acting on pyruvate or acetyl-CoA. This result is confirmed by radioactive isotopic, spectrophotometric and gas-chromatographic methods.     

Virulence factors of Entamoeba histolytica.

Recent studies have increased our knowledge of Entamoeba histolytica cell biology and gene regulation. In the ameba, dominant-negative mutations in the Gal/GalNAc lectin affect adhesion and cytolysis, whereas mutations in meromyosin affect cytoskeletal function. Studying these mutant proteins has improved our understanding of the role of these proteins in E. histolytica virulence. The characterization of the CP5 cysteine protease and the induction of apoptosis in host target cells has led to a better comprehension of the mechanisms by which trophozoites can lyse target cells.