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T F Jackson

Publications and source records attributed to T F Jackson.

At least 37 records · Page 2Linked to original sources
PubMed1992

Serodiagnosis of invasive amebiasis using a recombinant Entamoeba histolytica antigen-based ELISA.

Amebic serology remains a critical component in the diagnosis of invasive amebiasis. A recombinant serine rich-Entamoeba histolytica-protein/maltose binding protein (SREHP/MBP) fusion protein was evaluated as the target antigen in an ELISA test to detect acute invasive amebiasis. Retrospective analysis of 65 serum samples from patients with amebic liver abscess and 40 asymptomatic control patients showed the SREHP/MBP ELISA test had a sensitivity of 74% and specificity of only 55% for the diagnosis of amebic liver abscess. This study was repeated under identical conditions using a purified recombinant SREHP cleaved from the MBP component. The purified recombinant SREHP-based ELISA had a sensitivity of 79% and specificity of 87%. Our data suggest a purified recombinant SREHP-based ELISA could prove useful in the serodiagnosis of acute invasive amebiasis.

Animals↗
PubMed1991

Serodiagnosis of invasive amebiasis using a recombinant Entamoeba histolytica protein.

One hundred eight serum samples from 106 patients were examined by Western blot analysis for the presence of antibodies to a recombinant fusion protein containing the sequence of the newly described serine-rich Entamoeba histolytica protein (SREHP). Among patients with invasive amebiasis from Durban, Republic of South Africa; San Diego, Calif; Mexico City, Mexico; and St Louis, Mo, 53 (82%) of 65 had antibodies to SREHP. In contrast, only one patient (2%) of 43 without acute invasive amebiasis had antibodies to SREHP. The predictive value of a positive test for anti-SREHP antibodies in the detection of acute invasive amebiasis was most marked when analyzed in the patients from Durban, where 11 (92%) of 12 patients who were seropositive for SREHP had acute invasive amebiasis vs 17 (65%) of 26 patients who had a positive serologic diagnosis as determined by agar gel diffusion. The use of a serologic test based on the recombinant SREHP fusion protein may be a useful adjunct to the diagnosis of acute invasive amebiasis in endemic regions.

Amebiasis↗
PubMed1991

A comparative assessment of commonly employed staining procedures for the diagnosis of cryptosporidiosis.

Following an increase in the number of reports of Cryptosporidium infections and the problems encountered in detecting these organisms in faecal smears, a comparative assessment of a modification of the Sheather's flotation technique and other commonly employed staining procedures proved the modified Sheather's technique to be most useful in identifying Cryptosporidium oocysts in diarrhoeal stools. This technique not only detected the parasite in the highest number of stools but also proved to be cost-effective and the least time-consuming. Other staining techniques assessed were the modified Ziehl-Neelsen, safranin-methylene blue and auramine-phenol fluorescence. Both the modified Ziehl-Neelsen and the auramine-phenol fluorescence procedures produced nonspecific staining, while the safranin-methylene blue method was found to be the least sensitive technique.

Animals↗
PubMed1991

Cryptosporidium infections in children in Durban. Seasonal variation, age distribution and disease status.

One hundred and eleven of 1,229 children (9%) aged less than 10 years admitted to King Edward VIII Hospital, Durban, with gastro-enteritis over a period of 1 year were found to harbour Cryptosporidium. Of these, 96 (89.7%) were less than 2 years of age. Cryptosporidium was the only potential pathogen identified in 80 of these patients (6.5%). The prevalence in paediatric patients without gastro-enteritis was 2.4% (3/124). During the study period Cryptosporidium infections were significantly more prevalent during the high rainfall season (P = 0.03).

Age Factors↗
PubMed1991

Isolation of a strain-specific Entamoeba histolytica cDNA clone.

Entamoeba histolytica is an intestinal parasite causing significant morbidity and mortality worldwide. More tools are needed to understand the epidemiology and molecular pathogenesis of amebiasis. A cDNA library was constructed by using poly(A)+ RNA isolated from an axenic strain of E. histolytica, HM1:IMSS, which expresses a pathogenic isoenzyme pattern (zymodeme). Differential screening of the library yielded a strain-specific 3' polyadenylated cDNA clone, C2, possessing nine 26-nucleotide tandem repeats. RNA and DNA transfer blot analysis of four axenic strains of E. histolytica possessing the same pathogenic zymodeme revealed that the gene is present and expressed in pathogenic E. histolytica HM1:IMSS and 200:NIH but is not present in pathogenic strains HK-9 and Rahman. In addition, Southern blot analysis using the C2 clone showed heterogeneity of genomic organization between HM1:IMSS and 200:NIH. DNA dot blot hybridization analysis demonstrated that cDNA clone C2 was also able to distinguish axenically cultured E. histolytica strains possessing pathogenic zymodemes from those possessing nonpathogenic zymodemes and could detect as few as 100 amebic trophozoites. We conclude that C2 is a strain-specific E. histolytica cDNA clone that, in conjunction with other E. histolytica-specific probes, could serve as a useful epidemiologic tool.

Animals↗
PubMed1990

Monoclonal antibodies directed against the galactose-binding lectin of Entamoeba histolytica enhance adherence.

The Entamoeba histolytica galactose-binding lectin is a surface glycoprotein composed of 170- and 35-kDa subunits. Inhibition of this lectin with galactose or anti-170 kDa subunit polyclonal antibody blocks amebic adherence to target cells and colonic mucin glycoproteins. We describe the properties of 10 mAb with specificity for the 170-kDa subunit. Based on competitive binding studies, six nonoverlapping antigenic determinants on the lectin were identified. The effect of the mAb on adherence of amebic trophozoites to both Chinese hamster ovary (CHO) cells and human colonic mucins was measured. Antilectin antibodies directed against epitopes 1 and 2 enhanced adherence, with the number of amebae having at least three adherent CHO cells increasing with the addition of epitope 1 mAb from 26 +/- 9 to 88 +/- 2% and the binding of colonic mucins increasing from 34 +/- 1 to 164 +/- 3 pg/10(5) amebae. Antibody-enhanced adherence remained 90 to 100% galactose inhibitable, occurred at 4 degrees C and was not Fc mediated. Univalent Fab fragments of epitope 1 mAb augmented mucin binding by 238% and CHO cell adherence by 338%. The binding of purified lectin to CHO cells was increased from 1.1 +/- 0.1 to 2.4 +/- 0.3 ng/10(3) CHO cells by mAb directed to epitope 1, demonstrating that enhanced adherence was due to direct activation of the lectin. mAb to epitope 3 bound to the lectin only upon its solubilization from the membrane and had no effect on adherence. Adherence to CHO cells and mucins was inhibited from 50 to 75% by mAb to epitopes 4 and 5; epitope 6 mAb inhibited amebic adherence to CHO cells but not mucins. The pooled sera from 10 patients with amebic liver abscess blocked the binding to the 170-kDa subunit of mAb directed to all six epitopes. Striking individual variations in the effects of immune sera on adherence were observed. Although the sera of all 44 South African patients with amebic liver abscess had high titer anti-lectin antibodies, 16 patients' sera significantly (more than 3 SEM) enhanced adherence whereas 25 patients' sera significantly inhibited adherence. Antilectin antibodies exert profound functional effects on the interaction of E. histolytica with target cells and human colonic mucins. Exploration of the clinical consequences of adherence-enhancing and inhibitory antibody responses may give insight into the role of antilectin antibodies in immunity to invasive amebiasis.

Amebiasis↗
PubMed1990

Association of serum antibodies to adherence lectin with invasive amebiasis and asymptomatic infection with pathogenic Entamoeba histolytica.

The recognition of the Entamoeba histolytica galactose-inhibitable adherence lectin by antibodies was studied using sera obtained from subjects in South Africa with an amebic liver abscess or asymptomatically pathogenic or nonpathogenic E. histolytica infection and from uninfected regional controls. In addition, sera from healthy American controls or Americans known to be infected with other parasites were studied. Of the 95 sera containing antibodies to total parasite protein, 95% demonstrated antibodies to the 170-kDa heavy subunit but not to the 35-kDa light lectin subunit. All sera (n = 253) were tested by ELISA for antibodies to lectin: 99% from liver abscess patients and all 4 from individuals asymptomatically infected with pathogenic E. histolytica were positive; all from the 40 healthy American controls and the 29 infected with other parasites were negative (P less than .01). The prevalence of serum anti-lectin antibodies was identical (25%) in asymptomatic South Africans with either a nonpathogenic infection or a negative stool culture for E. histolytica. Thus, the presence of serum antibodies to lectin seems to indicate current or prior invasive amebiasis or asymptomatic intestinal infection with pathogenic E. histolytica.

Amebiasis↗
PubMed1990

Pathogenic and nonpathogenic strains of Entamoeba histolytica can be differentiated by monoclonal antibodies to the galactose-specific adherence lectin.

Entamoeba histolytica infection results in either asymptomatic colonization or invasive colitis and liver abscess. E. histolytica isolates from patients with invasive disease have characteristic isoenzyme profiles (pathogenic zymodemes), suggesting a role for parasite factors in determining the severity of infection. A galactose-specific cell surface lectin from a pathogenic zymodeme was shown to mediate in vitro adherence to human colonic mucins and contact-dependent killing of target cells. Six nonoverlapping antigenic determinants were identified on the 170-kilodalton heavy subunit of the pathogenic lectin. Anti-lectin monoclonal antibodies (MAb) directed against epitopes 1 and 2 enhanced adherence whereas MAb to epitopes 3 through 6 either inhibited or had no effect on adherence. We tested 50 pathogenic and nonpathogenic strains for reactivity to these anti-lectin MAb by radioimmunoassay. MAb to epitopes 1 through 6 reacted in the radioimmunoassay with all 16 pathogenic zymodeme strains tested. In contrast, only MAb to epitopes 1 and 2 bound to the lectin from nonpathogenic strains. Western immunoblots with anti-lectin antibodies showed that the 170-kilodalton heavy subunit was present in the nonpathogenic amebae. Adherence of the nonpathogenic SAW 760 strain to human erythrocytes was enhanced by MAb to epitope 1 and blocked by galactose, confirming the presence of a functionally active lectin. A lectin radioimmunoassay based on MAb to epitopes 1 and 3 proved to be a simple and rapid method to distinguish pathogenic from nonpathogenic amebae in culture. Further exploration of the functional consequences of the antigenic differences demonstrated for the lectin may lead to a better understanding of its role in pathogenesis.

Animals↗
PubMed1990

Detection of Entamoeba histolytica immunoglobulins G and M to plasma membrane antigen by enzyme-linked immunosorbent assay.

Sixty-one serum specimens from 22 patients with clinically diagnosed amoebic liver abscess (ALA), 10 hospitalized patients with a variety of diseases other than amoebiasis, 12 normal healthy controls, and 17 subjects from an amoebiasis-endemic area were assayed by enzyme-linked immunosorbent assay (ELISA). The plasma membrane fraction of axenic cultures of Entamoeba histolytica HK9 separated from other subcellular fractions by differential centrifugation was used as the antigen to detect specific immunoglobulin G (IgG) and IgM antibodies. Using a single serum dilution of 1/100 and optical densities at 492 nm of 0.200 and 0.250 as the cutoff values for the IgM and IgG ELISAs, their respective sensitivities in 22 ALA patients were 91% (20 of 22) and 95% (21 of 22). In 22 patients (10 hospitalized and 12 normal healthy controls), the specificities of the IgM and IgG ELISAs were 95% (21 of 22) and 91% (20 of 22), respectively. All five asymptomatic carriers of pathogenic E. histolytica were seropositive by the IgG ELISA and the amoebic gel diffusion test (AGDT). The AGDT was positive for three of six culture-negative controls, while the IgG ELISA was positive for all six. For six asymptomatic carriers of nonpathogenic zymodemes, the AGDT was positive for two, and the IgG ELISA was positive for three. There was an excellent correlation (r = 0.96) between the IgG ELISA and the AGDT. Only one of six culture-negative controls, none of the asymptomatic carriers of pathogenic E. histolytica, and one of six carriers of nonpathogenic E. histolytica were seropositive by the IgM ELISA, thus highlighting the specificity of the IgM ELISA in the diagnosis of ALA. It is believed that the use of plasma membrane fractions has improved the diagnostic potential of the IgM ELISA.

Animals↗
PubMed1990

Entamoeba histolytica: naturally occurring infections in baboons.

Entamoeba chattoni frequently occurs as an intestinal infection in non-human primates. It has been isolated from both wild and captive animals. Morphologically this amoeba resembles E. histolytica. E. histolytica has also been isolated from non-human primates on a number of occasions but these isolations have been from captive animals. In recent years identification of E. histolytica has been enhanced by the introduction of iso-enzyme electrophoresis methods. This technology has been widely applied to amoebae isolated from humans from many parts of the world. Limited work using iso-enzyme electrophoresis on non-human primates in captivity has confirmed the earlier parasitological studies mentioned above as both E. histolytica and E. chattoni were isolated; furthermore zymodeme (strain or species based on the characteristic iso-enzyme electrophoretic pattern) analysis demonstrated that these two organisms could be readily distinguished by this method. The importance of establishing whether E. histolytica occurs in wild primates as a true zoonosis was emphasised by these studies, particularly from the point of view of potential disease transmission. A study of baboons (Papio ursinus) in South African game reserves has been initiated and three surveys have been conducted in the Kruger National Park; these baboons are considered to live in isolation from humans. Of the 210 isolations attempted 65-75% yielded E. chattoni while E. histolytica were cultured from the faeces of 1-3% of the baboons. Although the prevalence rates varied in each of the 3 study locations both species of amoeba were found in baboons from all of them. All the E. histolytica isolated thus far have been non-pathogenic zymodemes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Evaluation of an enzyme-linked immunosorbent assay in the serodiagnosis of amoebic liver abscess.

An indirect enzyme-linked immunosorbent assay (ELISA) was evaluated for the detection of anti-amoebic IgG and IgM antibodies to assess its value in distinguishing past from current infection in invasive amoebiasis, particularly in amoebic liver abscess (ALA) patients. Using sera from 295 individuals, the ELISA was also compared with the amoebic gel diffusion (AGD) test. In 100 patients the IgG-ELISA at a single test dilution of 1/6,400 had a sensitivity of 99% for clinically diagnosed ALA. In these same patients the IgM-ELISA at a single dilution of 1/400, had a sensitivity of 64% and a specificity of 97.9%. No cross-reactions were observed in sera from patients with collagen vascular disease. In 121 patients without clinical invasive amoebiasis, 8 were AGD-positive and 12 were IgG-ELISA-positive, giving the latter assay a specificity of 91.7%. This is thought to be due to past infection with Entamoeba histolytica. In symptomless carriers of pathogenic zymodemes, 10/11 were seropositive by the IgG-ELISA and 11/11 by the AGD test. There was an excellent correlation between the IgG-ELISA and the AGD test (r = 0.99). The IgG-ELISA is a sensitive, specific, simple and rapid test. It has the clinical advantage that results are obtainable 2 1/2 hours after receipt of the specimen, compared with the 24-48 hours required for the AGD test. The prompt availability of IgG-ELISA results could prove advantageous for implementation of early therapy. The IgM-ELISA was not found to be sensitive enough to be used as an index of active amoebic infection.

Enzyme-Linked Immunosorbent Assay↗
PubMed1988

Biological evidence of genetic exchange in Entamoeba histolytica.

The demonstration of a new zymodeme of Entamoeba histolytica produced in culture from cloned isolates suggested possible genetic exchange in this parasite. We have attempted to substantiate that finding by using rats as biological hosts. Clones were made from 3 separate isolates of E. histolytica, each established in culture from liver pus or faeces. After enzyme characterization these clones, of zymodemes II, XIV and XIX, were paired in each of the 3 possible combinations and the mixtures injected into the caecum of rats. Clones of new or hybrid zymodemes were produced as well as the original parents, with one exception, the mixture of XIV and XIX, from which only one of the parents was recovered. The hybrids produced included zymodeme XX, observed previously, and zymodeme XI, a naturally occurring zymodeme that has in the past been recovered only from subjects with invasive amoebiasis.

Animals↗
PubMed1987

A longitudinal study of asymptomatic carriers of pathogenic zymodemes of Entamoeba histolytica.

The value of treating subjects who pass Entamoeba histolytica cysts in their faeces is currently in question. In the endemic Durban area iso-enzyme electrophoresis of E. histolytica isolates indicated that 1% of the asymptomatic population are infected with pathogenic zymodemes. The outcome of these potentially invasive infections was established by means of a longitudinal survey. All subjects had strongly positive serological responses--10% of them developed amoebic colitis while the rest remained asymptomatic and spontaneous cure occurred within 1 year. Infections with pathogenic zymodemes occurred in family units and closely related individuals. Since they were cyst passers, good evidence was obtained for the existence of a carrier state for pathogenic E. histolytica. The value of sero-epidemiological surveys in determining the prevalence of pathogenic E. histolytica in a community was highlighted. Treatment of asymptomatic carriers with pathogenic zymodemes of E. histolytica is as essential as treatment of patients with invasive amoebiasis, and in both cases all contacts of affected individuals must also be treated if they are found to harbour pathogenic zymodemes of E. histolytica in order to ensure control of disease transmission.

Adolescent↗
PubMed1985

A comparative trial of metronidazole v. tinidazole in the treatment of amoebic liver abscess.

Forty-eight patients were treated for amoebic liver abscess, 27 with metronidazole (Flagyl; Maybaker) and 21 with tinidazole (Fasigyn; Pfizer). Although 4 patients treated with tinidazole and 2 treated with metronidazole required a second course of therapy, both drugs were highly effective and resulted in rapid clinical improvement. The only side-effect was oral candidiasis, which developed in 2 patients in each drug group.

Adult↗
PubMed1985

Amoebic isoenzymes.

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Entamoeba histolytica↗