Hyperimmunoglobulinemia E in the absence of atopy and filarial infection: the Huaorani of Ecuador.

Hyperimmunoglobulinemia E (HIGE) is associated with various conditions such as atopy, dermatitis, hypersensitivity reactions, and certain parasitic infections. In the course of vaccination initiatives in the province of Napo, eastern Ecuador, blood samples were collected from one of the two remaining rural subgroups of Huaorani Indians who in 1979 were reported to have the world's highest concentrations of IgE. One subgroup of Huaorani, the Dicaron, lives in a protected Amazonian region which has reportedly suffered from extensive pollution after petroleum industry exploration. Plasma was collected from 31 members of the Dicaron (age range 15-75 years), eight non-Dicaron Huaorani, and 16 Quichua Indians from the same province, and tested for IgE, IgG, IgM, IgA, and immunoglobulin allotypes. Subjects were examined for evidence of filariasis, a group of parasitic diseases associated with HIGE. Mean IgE concentration in the Dicaron was measured by CAP ELISA at 11,850 IU/mL (range 5000-33,000) while IgA and IgM concentrations were within normal limits compared to North American controls. IgG levels were slightly elevated and there was no evidence of filariasis. Compared to the Quichua and non-Dicaron Huaorani, two other Amerindian tribes in the Ecuadorian Amazon, the highest concentrations of IgE were recorded from the Dicaron who live within the allegedly polluted section of the Amazon. We conclude that an unexplained HIGE syndrome exists among only one subgroup of Huaorani, the Dicaron. Other eastern Ecuadorian Amerindians, such as the Quichua and resettled Huaorani, have IgE concentrations expected in a population with intestinal helminthiasis. Environmental factors cannot be excluded as the cause of HIGE in the Dicaron.

Immunoglobulin allotypes among the Bicolanos of Sorsogon province, Luzon, Philippines: implications of phenotypes for filariasis.

Immunoglobulin (Ig) allotypes are polymorphic genetic systems that show distinct racial arrays, thus making them powerful tools for studies of genetic admixture and biological relationships. In the province of Sorsogon, southern Luzon, Philippines, allotyping was completed on 252 persons residing in two neighboring villages. The people demonstrated 14 GM and 3 KM phenotypes. The frequency of homozygous KM3, KM1 and heterozygous KM1,3 was identical in these villages; however, half of the GM phenotypes present in one village were significantly less frequent in the other village. The frequency of KM and GM haplotypes was different from those reported in Filipino aboriginal groups, but similar to a population on Samar Island, the only other Filipino group for which Ig allotype data exist. Variability in the prevalence of parasitic disease such as lymphatic filariasis may in part reflect differences in genetic susceptibility, resulting from allotypic heterogeneity between villages.

Anti-KS: identification of autoantibodies to asparaginyl-transfer RNA synthetase associated with interstitial lung disease.

Autoantibodies to five of the aminoacyl-transfer RNA (tRNA) synthetases have been described, and each is associated with a syndrome of inflammatory myopathy with interstitial lung disease (ILD) and arthritis. Serum KS, from a patient with ILD and inflammatory arthritis without evidence of myositis, immunoprecipitated a tRNA that was distinct from that precipitated by any described anti-synthetase or other reported tRNA-related Abs, along with a protein of 65 kDa. KS serum and IgG fraction each showed significant (88%) inhibition of asparaginyl-tRNA synthetase (AsnRS) activity, but not of any of the other 19 aminoacyl-tRNA synthetase activities. Among 884 patients with connective tissue diseases tested, only two other sera were found to immunoprecipitate tRNAs and proteins of identical gel mobility. These two and KS showed identical immunodiffusion lines using HeLa cell extract. The new sera significantly inhibited AsnRS without significant effects on other synthetases tested. Both patients had ILD but neither had evidence of myositis. These data strongly suggest that these three sera have autoantibodies to AsnRS, representing a sixth anti-synthetase. Anti-KS was more closely associated with ILD than with myositis. Further study of this Abs might prove useful in dissecting the stimuli responsible for the genesis of anti-synthetase autoantibodies.

Immunoglobulin allotypes in Ecuadorian Cayapa Indians.

Indigenous Indian groups comprise approximately 20% of Ecuador's population, the third largest percentage in all of Central or South America, yet immunogenetic data on these groups are lacking in the literature. In the course of population migration studies, sera collected from 65 Ecuadorians living in the northern province of Esmeraldas were typed for six GM and two KM markers. The study population consisted of 47 Cayapa Indians and 18 blacks of African origin, descendants of slaves imported into the area during the seventeenth century. The Cayapa demonstrated three GM phenotypes, two of which are common to other South American Indian tribes. The frequency of KM1 positive Cayapa Indians (63%) is similar to other South American Indian tribes, but is significantly greater than the Huaorani of eastern Ecuador (2%), the only other Ecuadorian Indian group for whom limited immunoglobulin allotype data are available (chi 2 = 35.8, P < 0.0001).

Antibody responses to Onchocerca volvulus in Ecuadorian Indians and blacks.

In Ecuador, previous epidemiological studies of onchocerciasis affecting a population consisting of Blacks of African origin and Amerindians of the Chachi tribe, have demonstrated different frequencies of clinical disease manifestations in each racial group. To explore the possibility of differential antibody responses to O. volvulus antigens in this population, a subset of sera from these two groups of Ecuadorians with onchocerciasis was analyzed for antibody to O. volvulus antigens using ELISA and Western blot techniques. Significantly higher levels of immunoglobulin against O. volvulus were demonstrated in Chachi Indians compared to native Blacks of African origin: total Ig (p = 0.049), IgG (P = 0.002) IgG4 (p = 0.019) and IgA (p = 0.035). Western blot analysis of sera from 41 persons demonstrated more intense and more frequent recognition of low molecular weight parasite antigens in the Chachi compared to a similar group of male and female Ecuadorian Blacks. These data suggest that the intensity of antibody responses to O. volvulus antigens may reflect the influence of racial factors in the pathogenesis of onchocerciasis in Ecuador.

Characterization of a variant tandem repeat from Sudanese Onchocerca volvulus.

Strategies for detection and control of onchocerciasis in Africa have included identification of DNA probes and PCR target sequences for sensitive and specific detection of parasites. To evaluate the applicability of published PCR and DNA probe based methods for the study of onchocerciasis in Sudan, we collected adult O. volvulus from geographically distinct regions of Sudan (700 miles apart), Abu Hamed (northern desert) and Raja (southwestern savannah), and we examined the similarities between Sudanese O. volvulus repeats and published versions of the repeat from West African O. volvulus. Amplification of DNA extracted from the Raja O. volvulus strain predictably generated a ladder of products, multiples of the base 150 bp repeat, as has been reported from West Africa. However, amplification of DNA from the Abu Hamed O. volvulus isolate resulted in a series of doublets. The unexpected DNA fragments thus amplified differed in size from the base 150 bp unit by approximately 50 base pairs and was most clearly visualized at 150-200 base pairs. DNA sequence analysis of the amplified repeats in the isolate of O. volvulus from Abu Hamed revealed a variant of the 150 bp tandem repeat which contained an extra 49 bp. The additional 49 bp contained two short repeats of 21 bp and 10 bp, corresponding to bases 99-119 and 128-137 respectively, of the known 150 bp O. volvulus repeat. This work demonstrates a variant of the O. volvulus 150 bp tandem repeat, which easily distinguishes Raja and Abu Hamed isolates of O. volvulus, and which has potential value for differentiating Abu Hamed strains of O. volvulus from other strains in East Africa.

Identification of related DNA sequences in Borrelia burgdorferi and two strains of Leptospira interrogans by using polymerase chain reaction.

The suitability of a polymerase chain reaction assay for Borrelia burgdorferi in epidemiological studies of infected tick populations was evaluated by using 28 strains of Leptospira interrogans and lysates of fixed adult Ixodes tick tissues. Two false positives representing leptospires were differentiated from B. burgdorferi by using an oligonucleotide probe.

Surface area calculation for Onchocerca volvulus microfilariae.

The pathogenesis of tissue damage in filarial infections involves the generation of inflammatory mediators by interaction of parasite surfaces with the host immune response. In the presence of a heavy parasite burden, the tremendous potential for development of adverse local or systemic inflammatory responses may be appreciated by knowledge of the cumulative surface area represented by circulating microfilariae. The approximate surface area of a single microfilariae of Onchocerca volvulus was calculated using mathematical formulae for standard geometrical shapes and values derived from serial measurements of electron micrographs. Numerical values for the surface area represented by large numbers of microfilariae are given and compared to surface areas of the host which can harbor many millions of microfilariae.

Abdominal sparganosis in Ecuador: a case report.

Sparganosis, infection with plerocercoids of the pseudophyllidean tapeworm Spirometra, rarely has been described in Ecuador. We report the details of a human case of sparganosis identified serendipitously in the course of an abdominal hernia repair. The parasite was found moving freely upon the external oblique fascia adjacent to the site of a direct abdominal hernia. The organism was recovered intact, photographed while alive and preserved for subsequent detailed morphological studies. The presumed route of entry into this patient was percutaneous, after application of a poultice of snake flesh to the site of a painful abdominal hernia. The literature on sparganosis in South America is reviewed. This is the second case of human sparganosis reported from Ecuador.

Onchocerca volvulus recombinant antigen: physical characterization and clinical correlates with serum reactivity.

An Onchocerca volvulus cDNA clone expressing epitopes found in adult and larval parasites, designated lambda RAL-2, was derived from a 1,000-base message present in adult O. volvulus, which encodes a protein with an apparent molecular weight of 17,000. This protein does not appear to be extensively posttranslationally modified. Serum samples from 52 individuals exposed to O. volvulus were examined for antibodies recognizing the lambda RAL-2 recombinant antigen; 77% produced such antibodies. In addition, individuals producing antibodies recognizing the recombinant antigen were significantly less likely to develop some aspects of ocular pathology associated with O. volvulus infection than were individuals who did not do so. These results suggest that recombinant antigens such as that produced by lambda RAL-2 may be useful in attempts to understand the mechanism of O. volvulus-induced ocular pathology.

Molecular epidemiology of OHIO-1 beta-lactamase.

A total of 31 plasmids, all bearing a gene that encodes a novel, plasmid-mediated Richmond-Sykes class III beta-lactamase designated OHIO-1 and a gene that encodes aminoglycoside 2"-adenyltransferase, have been collected from hospitals in Ohio. By using restriction endonuclease digestion and Southern hybridization, we were able to demonstrate that all these plasmids have a common genetic origin.

Severe onchocercal dermatitis in the Ethiopian border region of Sudan.

Onchocerca volvulus infection of the residents of two villages in the Upper Atbara River region of Sudan (63.4% prevalence) was found to be medically and socially significant with a remarkable degree of pruritus and dermal pathology present. The severest skin changes, which were often confined to certain areas of the body, were most common in the teenage group (13-19 years). Quantitation of the clinical changes showed that the most severe alterations were present in patients with the lower levels of microfilariae in the skin (0.1-6 mf mg-1) rather than in those with higher levels. Nodules containing adult O. volvulus were found in many individuals, and often in clusters in older patients. Major losses of vision due to O. volvulus were not found, although microfilariae were frequently seen in the cornea and anterior chamber. The commonest corneal change was punctate keratitis, and this was most frequently seen in the teenage group. Trachomatous changes of eyelids and corneas were also seen. The prevalence values of O. volvulus infection in the two villages were significantly increased when, in addition to microfilarial presence in skin biopsies, onchocercal signs such as punctate keratitis, nodule presence and microfilariae in the anterior chamber were taken into consideration. This illustrates the need to consider factors other than skin snip parasitology in obtaining a diagnosis. This study also emphasizes the degree of severity that onchodermatitis can reach and that low levels of dermal microfilariae can be accompanied by very extensive skin changes without any loss of vision. The necessity of considering the severity of skin changes when defining tolerable levels of onchocerciasis in a community is discussed.

Cell adherence to microfilariae of Onchocerca volvulus: a comparative study.

The conditions were examined for in vitro antibody-mediated adherence of granulocytes to microfilariae of Onchocera volvulus and Dirofilaria immitis. Reactivity in human sera from patients in endemic foci in Sudan was specific for O. volvulus and no reactions were observed with heterologous Onchocerca species or with Mansonella perstans. Microfilariae from skin, nodules or adult female worms were satisfactory targets for cell adherence, and the cells involved were almost exclusively eosinophils. The reaction was inhibited by indomethacin but not by nordihydroguaiaretic acid, an inhibitor of leukotriene production. Agents that slowed or stopped microfilarial motility (e.g. nifedipine, lidocaine, chloroquine) inhibited the reaction, probably by reducing target/cell contact. Ivermectin did not enhance the reaction, and in the absence of cells exerted only slight effects on the movement of microfilariae at higher concentrations (greater than 10 micrograms/ml). Antibody activity was labile, and did not persist well through freeze-thaw cycles. Some differences between homologous and heterologous mixtures (microfilariae/cells/serum) were seen but they could not be resolved satisfactorily. There were no apparent geographical differences between microfilariae from different foci in Sudan. In the D. immitis system neutrophils were the dominant cell type adhering to microfilariae, and the activity was stable to storage and freeze-thaw. No enhancement was detectable with diethylcarbamazine. Antibody activity was absorbable with microfilarial antigens and was reduced by agents that inhibited microfilarial motility. In dogs, adherence-mediating antibody was seen only in amicrofilaraemic animals with occult infection, and in only a minority of these sera. In humans the relationship to clinical findings was less clear, but patients with punctate keratitis were the most likely to have positive serum and were the most reactive in the assay. This system may therefore offer some insights into disease mechanisms in vivo, and its molecular mechanisms deserve further characterization.

Novel plasmid-mediated beta-lactamase in members of the family Enterobacteriaceae from Ohio.

Epidemiologic studies of plasmid-mediated resistance at the Cleveland Veterans Administration Medical Center revealed that related plasmids had disseminated among members of the family Enterobacteriaceae. We studied the beta-lactamases encoded by these plasmids in Escherichia coli C600 transformants or transconjugants. Substrate and inhibition profiles of the enzymes determined by two of these plasmids suggested an activity resembling TEM-1; however, isoelectric focusing revealed a pI of 7.0. These two plasmids were originally found in a Serratia marcescens (pDS076) and an Enterobacter cloacae (pDS075) strain isolated from the same sink in the medical intensive care unit and later, in an Enterobacter cloacae (pDS142 identical to pDS076) isolate colonizing a patient in the same unit. The plasmids also carried the aminoglycoside resistance determinant, 2"-aminoglycoside nucleotidyl transferase. A 2-kilobase AvaI restriction endonuclease digestion fragment of pSD075 known to carry the beta-lactamase determinant was used as a molecular probe. This probe did not recognize sequences of any plasmid-mediated beta-lactamase tested including the recently described determinants ROB-1, TLE-1, and OXA-4-7. A TEM-1 probe derived from the 0.7-kilobase PstI-EcoRI fragment of pBR322 failed to recognize the new beta-lactamase gene. Four additional Enterobacter cloacae and two Enterobacter aerogenes strains isolated in Columbus, Ohio, have been shown to produce a pI 7.0 beta-lactamase and to carry plasmids recognized by the 2-kilobase probe. These data suggest dissemination of a novel plasmid-mediated beta-lactamase among members of the family Enterobacteriaceae in Ohio and demonstrate the development and utility of a molecular probe for the new determinant. We suggest that the novel beta-lactamase be named OHIO-1.

Pentamidine: a review.

Pentamidine, recently released for clinical use, is effective in therapy for the hemolymphatic stage of Gambian trypanosomiasis, antimony-resistant leishmaniasis, and Pneumocystis carinii pneumonia. The mechanism of action is unclear and may differ for different organisms. Trypanosomes actively transport pentamidine intracellularly, and the drug may then interfere with DNA biosynthetics. However, pentamidine appears to kill nonreplicating P. carinii. The mechanism of killing is unexplained. The pharmacokinetics of pentamidine has been incompletely studied in humans. The estimated volume of distribution is 3 liters/kg. Levels in plasma of pentamidine range from 0.3-1.4 microgram/ml after standard 4 mg/kg dosing, with no appreciable increase in drug levels on successive dosing and no correlation between levels and creatinine clearance or adverse reactions. The drug appears to be concentrated in the kidney and excreted in the urine, with levels detectable six to eight weeks after cessation of therapy. Immediate adverse reactions have included hypotension, nausea, and vomiting. Local pain or abscess formation at an injection site, mild azotemia, leukopenia, abnormal findings from liver function tests, and hypoglycemia may also occur.