The monocyte locomotion inhibitory factor produced by Entamoeba histolytica inhibits induced nitric oxide production in human leukocytes.

The monocyte locomotion inhibitory factor, an anti-inflammatory pentapeptide produced by Entamoeba histolytica, inhibits the in vitro production of nitric oxide induced by cytokines (INF-gamma, TNF-alpha) or PMA in human leukocytes. This can be added to the other previously reported functional effects of this factor, such as the inhibition of monocyte locomotion and the synthesis of reactive oxygen intermediates in both monocytes and neutrophils. The decreased nitric oxide production may interfere with the killing of amebas by neutrophils in the early invasive stages of amebiasis, when oxidative mechanisms are used [reactive oxygen and nitrogen intermediates either individually or synergistically via peroxynitrite (ONOO(-))], and in the advanced stages, when both non-oxidative and oxidative (including nitric oxide) mechanisms are employed by macrophages. Diminished nitric oxide production by leukocytes may also contribute to the paucity of late inflammatory components in amebic abscess of the liver and other amebic lesions.

A novel anti-inflammatory oligopeptide produced by Entamoeba histolytica.

The monocyte locomotion inhibitory factor (MLIF), a heat-stable oligopeptide found in the supernatant fluid of Entamoeba histolytica axenic cultures was isolated by ultra-filtration, gel-sieve chromatography and high powered liquid chromatography (HPLC), and its primary structure (Met-Gln-Cys-Asn-Ser) established by Edman sequencing and mass-spectrometry (MS). A synthetic peptide had the same selective anti-inflammatory features as the native material in comparable concentrations: in vitro inhibition of the locomotion in human peripheral blood monocytes, and of the respiratory burst in the same cells and in human neutrophil polymorphonuclear leucocytes; and in vivo depression of delayed hypersensitivity skin reactions to dinitrochlorobenzene in guinea pigs. This oligopeptide is apparently synthesized by the ameba as suggested by [(35)S]-Cys and Met incorporation, probably as part of a larger molecule, from which it is cleaved by proteolysis. The full sequence was not found in the 431 available E. histolytica protein sequences. The factor may contribute to the unexpected paucity of the late inflammatory reaction found in advanced invasive amebiasis and, perhaps in consequence, to the regeneration without scarring (restitutio ad integrum) of the affected organs that is observed following successful treatment of this disease

Does the eosinophil have a protective role in amebiasis?

While normal human eosinophils are destroyed in vitro by virulent Entamoeba histolytica, notwithstanding the presence of antibodies and complement, activated eosinophils promptly destroy the parasite although dying also at the end of the process. To study the possible in vivo participation of eosinophils in invasive amebiasis, we compared the induction of experimental amebic abscess of the liver (AAL) in gerbils (Meriones unguiculatus) previously made eosinophilic through Toxocara canis antigen injection and in normal control gerbils. After intraportal inoculation of 10(5) ameba trophozoites (6 and 24 hr), the ratio of gerbils with AAL, as well as the number and size of the microabscesses was comparable in eosinophilic and control gerbils. However, at 96 hr the number and size of the microabscesses were significantly smaller (p < 0.05) in eosinophilic gerbils. On the other hand the actuarial AAL survival curve up to 45 days post-amebic inoculation was significantly (p < 0.05) shifted to the right in controls. These results suggest that antigen-induced eosinophilia may exert a protective effect against AAL in gerbils.

Increased frequency of HLA-DR3 in Mexican mestizo pediatric patients with amebic liver abscess (ALA).

Mexican mestizo pediatric patients with ALA revealed a significantly increased frequency of HLA-DR3 alone, or in its haplotype form HLA-A2, DR3, which confirms our previous observation in adult patients with ALA in the same ethnic group. However, the relative increase in these HLA specificities in pediatric ALA patients when compared to adult patients was not statistically significant, and calls for an enlargement of the population studied.

Increased frequency of HLA-DR3 and complotype SC01 in Mexican mestizo patients with amoebic abscess of the liver.

Our preliminary study (31 patients) of HLA frequencies and amoebic abscess of the liver (AAL) in Mexican mestizos was extended to include 110 patients with this condition. The previously found increase in HLA-B16 was not confirmed, but the frequency of HLA-DR3 was again found significantly increased in patients with AAL when compared to the normal, ethnically matched control population, both in its isolated (35.5% vs 12.7%) and in the HLA-A2, DR3 haplotypic version (20.9% vs 4.5%). Moreover, seven of the 17 HLA specificities that were found to be individually different at P (yet not at PC) level in patients with AAL when compared to the control population, were actually HLA-DR3 containing haplotypes. HLA-DR3 may thus encode a risk factor(s) for AAL, at least in the Mexican mestizo population. Furthermore a significant increase in the complotype SC01 and its haplotypic version SC01, DR4 were identified in 45 non selected patients with AAL when compared to normal controls (31.1% vs 6.7% and 17.8% vs 0% respectively). Even though a relationship between allelic forms of complement components and their function has not been fully established, this complotype could represent a risk factor as well, since complement appears to play a role in host defence against amoebic invasion. Finally, no extended haplotype preference was found in these AAL patients.

[Monocyte locomotion inhibiting factor (MLIF) produced by E. histolytica induces an increase of cAMP in human monocytes]. / El factor inhibidor de la locomoción de monocitos (FILM) producido por E. histolytica induce aumento del AMPc en los monocitos humanos.

The supernatant fluid of axenically grown E. histolytica contains a factor (MLIF) which inhibits the locomotion of human monocytes (including chemotaxis) without affecting that of human polymorphonuclear leucocytes. Locomotion, like other cellular functions, is modulated by changes in intracellular cAMP and cGMP. The consensus--with some exceptions--is that while rises in cGMP accompany locomotion, an increase in cAMP (without a concomitant fall in -cGMP) occurs with inhibition of cellular movement. We measured by radioimmunoassay the cAMP concentration of human monocytes exposed to inhibitory concentrations of MLIF. A significant (p less than 0.005) rise in monocyte cAMP was found, comparable to that observed with the use of forskolin, a well known cAMP stimulator. The control studies using plain axenic medium, not only failed to reveal any rise in cAMP but disclosed a small, yet not significant drop in intracellular cAMP. These results suggest that MLIF (like other locomotion inhibitors, i.e. prostaglandins E1, A1 and isoproterenol) produces a significant increase in intracellular monocyte cAMP. This modification in intracellular signals may contribute to the inhibition in monocyte locomotion, an event during which an increase in pericentriolar microtubules has also been observed.

[HLA antigens, complement types, and amebic liver abscess in Mexican mestizos]. / HLA, complotipos y absceso hepático amibiano en mestizos mexicanos.

The association between parasitic diseases, and antigens of the major histocompatibility complex in man (HLA) has been poorly studied. The only study of the association between the HLA system and amebic abscess of the liver was performed by our group, and revealed a significant increase in HLA-BI6 and HLA-DR3 in patients with amebic abscess of the liver. The present study covered a larger number of patients and controls (110 of each) and it confirmed the association with HLA-DR3, it failed to confirm the association with HLA-BI6.

In vitro and in vivo effect of heparin, chondroitin, dextran and protamine on the virulence of pseudorabies virus (Suid herpesvirus 1).

The possible antiherpetic effect of three polyanions (heparin, chondroitin sulfate and dextran sulfate), as well as one polycation (protamine sulfate) was tested in vitro and in vivo against pseudorabies virus (Suid herpesvirus 1). The in vitro experiments revealed that heparin, dextran sulfate and protamine sulfate significantly reduced the number of lytic plaques. Chondroitin sulfate only caused a decrease in mean plaque size. Experiments in vivo disclosed that heparin injected subcutaneously before the experimental infection, was the only polyanion that protected mice against pseudorabies virus. Protamine sulfate had a paradoxic effect: whereas in vitro it reduced the number of lytic plaques, in vivo it increased the lethality of pseudorabies virus. Chondroitin sulfate and dextran sulfate did not modify the virulence of the virus in mice.

HLA antigens associated to amoebic abscess of the liver in Mexican mestizos.

Worldwide prevalence of amoebiasis is estimated at 4 x 10(8) cases/year, yet only one of about 300 individuals harbouring Entamoeba histolytica suffers tissue invasion and these cases are mostly concentrated in certain areas of Asia, Africa and Latin America. Patients with amoebic abscess of the liver (AAL) represent only a small fraction of that. These contrasting figures have been tentatively explained on the one hand through variations in sex, immunocompetence, nutritional and other socioeconomic features of the host, and on the other hand through differences in parasite virulence. In order to explore a possible association between the major histocompatibility complex (MHC) and AAL susceptibility, we studied the HLA profile in 31 Mexican mestizos with AAL and compared it to race and socioeconomically matched controls. Mexican mestizo patients with AAL revealed a significant increase in HLA-Bw16 and HLA-DR3 which could suggest an HLA-related susceptibility to liver invasion by E. histolytica.

HLA-B27 and ankylosing spondylitis in the Mexican Mestizo population.

Two previous surveys of ankylosing spondylitis (AS) in Mexican Mestizos found HLA-B27 frequencies of 68.6% and 78% and a relative risk (RR) of 37.05 and 120.88, respectively. We examined an additional group of Mexican Mestizos with AS and found an HLA-B27 frequency of 80.77% and a RR of 99.24. Our results are statistically comparable to the previous studies, and they suggest that the Mexican Mestizo is similar to the Spaniard in regards to AS and HLA-B27 association.

Efecto de la cicloheximida sobre la inhibicion de la quimiotaxia de los monocitos humanos provocada por productos de E. histolytica. / Effect of cycloheximide on inhibition of chemotaxis of human monocytes induced by products of E.histolytica

La cicloheximida es capaz de cancelar la inhibicion de la quimiotaxis de monocitos humanos provocada por sobrenadantes de cultivos axenicos de E. histolytica. Esto es consistente con el concepto de que la inhibicion ocurre al nivel celular y no por deactivacion de los atraentes. Este efecto ocurre a concentraciones de cicloheximida superiores a las necesarias para lograr una inhibicion de sintesis de proteina del 90 por ciento, por lo que el efecto cancelante de la inhibicion, o se debe a una proteina de metabolismo excepcional, u ocurre independientemente de la sintesis de proteinas