Immunological techniques in and for developing countries.
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Animals↗
Biomedical subjects
E P Wright
Publications and source records attributed to E P Wright.
Poorly differentiated carcinoma of unknown primary site: clinical usefulness of immunoperoxidase staining.
To assess the clinical utility of immunoperoxidase tumor-cell staining in patients with poorly differentiated carcinoma of unknown primary site, we performed a battery of stains on tumors from 87 patients treated between August 1978 and April 1983. Poorly differentiated carcinoma or poorly differentiated adenocarcinoma was diagnosed on the basis of light microscopic examination, and all patients were treated before the technology of immunoperoxidase staining was routinely used. Therefore, results of immunoperoxidase staining can be correlated with clinical outcome in this group of similarly treated patients with a long median follow-up. Immunoperoxidase staining confirmed the diagnosis of poorly differentiated carcinoma in 49 patients (56%) and yielded other diagnoses in 14 patients (16%): melanoma, eight; lymphoma, four; prostatic carcinoma, one; and yolk sac carcinoma, one. In 24 patients (28%) the immunoperoxidase staining pattern was inconclusive; electron microscopy was usually helpful in clarifying the diagnosis in these patients. Seventy-five patients (86%) received combination chemotherapy with a cisplatin-based regimen, and 24 patients (28%) had a complete response. Nine of these patients were later given specific diagnoses by immunoperoxidase staining (lymphoma, four; melanoma, four; yolk sac tumor, one). All patients with an immunoperoxidase diagnosis of lymphoma also had clinical features compatible with lymphoma and are long-term survivors. Patients with immunoperoxidase features suggesting melanoma were surprisingly responsive to chemotherapy, with three of seven complete responses and two long-term survivors. Patients with melanoma diagnosed by immunoperoxidase staining should not be excluded from a trial of cisplatin-based therapy. Immunoperoxidase staining is useful in the routine evaluation of metastatic poorly differentiated carcinoma of unknown primary site, as it can occasionally suggest the lineage of the tumor and have specific therapeutic implications.
Adenocarcinoma↗
Skin test responsiveness to a series of new tuberculins of children living in three Vietnamese cities.
A skin test survey was conducted among 1035 children aged 7-19 years living in three cities in Vietnam. Fifteen new tuberculins, including leprosin-A, were applied; an induration of 2 mm diameter or more was considered positive. Compared to some other tropical countries, low levels of sensitisation were recorded and remarkable regional differences were found. Positivity to any tuberculins (pooled data) among non-BCG-vaccinated children was significantly lower in Hanoi (13.1%) and HoChiMinh-City [HCMC] (15.5%) than in Nha Trang (25.7%) [p = 0.001 and p = 0.012, respectively]. The proportion of non-vaccinated children responding to Tuberculin ranged from 18.4% in Hanoi to 54.5% in Nha Trang. Leprosin-A elicited a response in 14.9% of the children in Nha Trang, but in very few of those in Hanoi (4.3%) or HCMC (3.0%). Thus, of the three cities studied, significant sensitisation to both M. tuberculosis and M. leprae was demonstrable only in Nha Trang. In Hanoi most of the response was to fast-growing species whilst in HCMC and Nha Trang it was mainly to slow-growing species. These results may account in part for the observed differences in the prevalence of tuberculosis and leprosy between the north and the south of Vietnam.
Adolescent↗
Prevention of cancer at the urothelial enteric anastomotic site.
An animal model of ureterosigmoidostomy was employed to elucidate the etiology of urinary intestinal anastomotic cancer. One hundred ninety-four rats were divided into five groups: vesicosigmoidostomy rats fed water without drug (controls); those fed ammonium chloride, or sodium bicarbonate, or ascorbic acid; and, finally, vesicosigmoid interposition rats. A significantly decreased incidence of cancer was evident only in the bowel interposition group (p less than 0.001). Ascorbic acid and altered urinary pH were not prophylactic. Possible mechanisms are discussed.
Anastomosis, Surgical↗
Primary meningococcal arthritis.
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Adolescent↗
Leishmania infection: surfaces and immunity.
Infections with Leishmania parasites are initiated by bites from infected sandflies; the injected promastigotes are attacked by phagocytic cells but succeed in entering cells of the macrophage family and surviving in them. The secrets of the success of the extracellular form in penetrating the host cell and of the intracellular form in surviving in a potentially hostile environment are yet to be unraveled. The infectivity of the extracellular promastigote is related to the expression on its surface of molecules that interact with the surface of the host cell. One of these molecules is the promastigote surface protease, or gp63, which is also a dominant surface antigen; this enzyme is thought to be involved in binding to the macrophage via the cell receptors for mannose and fucose and for the third component of complement. Another important surface component is the lipophosphoglycan, consisting of a series of phosphorylated disaccharides linked to a novel lipid anchor in the membrane. This is also released from the parasite surface and was earlier identified as a highly immunogenic antigen excreted into culture medium. It can activate complement and may in this way promote attachment of the parasite to the macrophage. Other surface structures include the acid phosphatase, a glyco-inositol phospholipid, another glycolipid, and membrane proteins of 80 and 17 kilodaltons. All of these may play a role in attachment of the promastigote to the macrophage host cell, as well as in the survival of the amastigote within the macrophage, perhaps by inhibiting the activities of destructive enzymes. The roles in infectivity of these components of the Leishmania surfaces and their interactions with the various receptors on macrophages are discussed. The immune responses induced by these and other parasite antigens during infections in humans and experimental animals are also described briefly, especially those responses that may contribute to protection from infection, or to diagnosis and epidemiology.
Animals↗
Leishmania donovani donovani antigens recognized by kala-azar patient sera and identification of cross-reacting antigens to Chagas' disease.
L. donovani antigens were analyzed by a direct agglutination test (DAT), by ELISA using intact promastigotes and by the SDS-PAGE immunoperoxidase technique (SGIP). Sera of Chagas' disease patients cross-reacted in the ELISA and SGIP but not in the DAT. Trypsin treatment of the parasites removed concanavalin A-binding sites but not epitopes for antibodies present in Chagas' disease and in leishmaniasis sera, as seen in the SGIP. Eight bands were revealed after incubation of the gel sections with kala-azar or Chagas' disease sera, three of which were common to both sera. The major antigenic component recognized by leishmaniasis sera was a glycoprotein of 57 kD, and the major cross-reacting protein recognized by Chagas' disease sera was a glycoprotein doublet of 71.5-68 kD.
Agglutination Tests↗
Morphological changes and serological reactions in cultured Leishmania donovani promastigotes.
Promastigotes of Leishmania donovani cultured in liquid media gradually changed from a majority of elongated forms through a predominance of intermediate spindle-shaped parasites to a majority of round forms, often aflagellate. Quantitative and qualitative differences among these subpopulations, separated on Percoll gradients, could be demonstrated when they were run on SDS-PAGE gels and stained from proteins or carbohydrates. When cultures containing a majority of one or other form were used as antigenic substrate in serological tests (ELISA and immunofluorescence) the older, round forms were more highly reactive with patients' sera. These forms may represent different phases of the parasite, related to function, and the density variations would provide a convenient method to separate them.
Animals↗
Poorly differentiated carcinoma of unknown primary site: correlation of light microscopic findings with response to cisplatin-based combination chemotherapy.
We have previously reported complete responses and long-term survival in patients with metastatic poorly differentiated carcinoma (PDC) of unknown primary site who received intensive cisplatin-containing chemotherapy regimens. We reviewed the light microscopic specimens from 113 patients with PDC in an attempt to identify common histopathologic features in the chemotherapy-responsive subgroup, and to rule out the presence of previously unrecognized germ cell tumors. Relatively few diagnoses more specific than PDC could be made. We could identify no histopathologic features by light microscopy that distinguished responsive from unresponsive neoplasms. Only one patient was found to have a previously unrecognized yolk sac carcinoma, and in five other patients the possibility of a germ cell neoplasm was considered in the differential diagnosis by at least one reviewer. The remaining tumors had no histologic features suggestive of germ cell neoplasms. Ninety-six patients had received combination chemotherapy (89 with cisplatin-containing regimens); 27 patients (28%) achieved complete remission, and 16 remain free of disease at a median of 65 months after completion of therapy. Patients with PDC of unknown primary site who are responsive to cisplatin-containing chemotherapy regimens cannot be reliably identified by light microscopy. At present, all such patients should be considered for an empiric trial of chemotherapy with cisplatin-based regimens, since cure is achievable in a minority.
Antineoplastic Combined Chemotherapy Protocols↗
Monoclonal antibody identification of Leishmania tropica from Sudanese mucosal leishmaniasis.
Monoclonal antibodies have been raised against a stock of Leishmania parasites which were isolated from a patient with mucocutaneous leishmaniasis in the Sudan. This stock did not react with monoclonal antibodies to L. donovani, and monoclonal antibodies raised to this stock were not reactive with stocks of L. donovani. They did react with stocks of L. tropica. The antibodies recognized antigens of high and of low molecular weights which were detectable in immunofluorescence on cultured promastigotes and on amastigotes cultured in macrophages. The taxonomy of the species responsible for the different clinical manifestations of infection with Leishmania is not yet clear; there has been for example discussion as to whether the different forms of the disease in the Sudan are caused by one or more species of parasite. Studies with highly specific monoclonal antibodies can help to clarify this picture.
Animals↗
Lipid laden macrophages in synovial fluid: a late finding in traumatic arthritis.
A 66-year-old man presented with arthritis of the right knee of 3 months' duration. Cytologic examination of synovial fluid (SF) showed numerous lipid laden macrophages. Radiographs and a nuclear scintigram established the presence of a fracture of the lateral tibial plateau. The diagnostic significance of lipid laden macrophages in SF is reviewed.
Aged↗
Serodiagnosis of Sudanese visceral and mucosal leishmaniasis: comparison of ELISA--immunofluorescence and indirect haemagglutination.
Two established immunodiagnostic techniques, immunofluorescence and indirect haemagglutination, were compared with ELISA (enzyme-linked immunosorbent assay) using intact promastigotes as antigen for the detection of specific antibodies against Leishmania in the serum of patients with visceral or mucosal leishmaniasis from the Sudan. The ELISA was found to be more sensitive and more specific than either of the other two tests.
Cross Reactions↗
Characterization of the humoral immune response in Sudanese leishmaniasis: specific antibody detected by class- and subclass-specific reagents.
Sera from Sudanese patients with either visceral or mucosal manifestations of infection with Leishmania were investigated to determine the class of antibodies against the causative parasite in an enzyme-linked immunosorbent assay (ELISA) with intact promastigotes as antigen. All patient sera had significantly more IgG and more antiparasite IgG than did control Sudanese or control Dutch sera; little or no IgM or IgA antiparasite antibody activity was detected. When sera specific for the subclasses of IgG were used to detect the anti-promastigote antibodies, these were found in IgG1- and IgG3-specific ELISA but not in those for IgG2 or IgG4.
Adolescent↗
Macrophage-induced procoagulation assay for the detection of defective macrophage activation in leprosy patients.
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Blood Coagulation Factors↗
Infection with parvovirus during pregnancy.
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Adult↗
ELISA using intact promastigotes for immunodiagnosis of kala-azar.
The commonly used diagnostic methods for visceral leishmaniasis (kala-azar) have many disadvantages. ELISA appears to be a more easily applied test for diagnosis, follow-up of treatment and epidemiology. The preparation of the ELISA antigen has given rise to difficulties in standardization since these are usually soluble. Intact cultured promastigotes have been used to develop an ELISA which is sensitive and specific while avoiding the extra handling which is a source of variability and lack of reproducibility. When tested on sera from kala-azar patients and patients with other tropical diseases, this ELISA was able to detect specific antibodies at very high serum dilutions. It is thus promising as a screening method applicable in the field.
Antibodies↗
Aortic intimal sarcoma with embolic metastases.
A 46-year-old woman died from massive bowel infarction. At autopsy, a primary sarcoma was found growing along the intimal surface of the aorta at the level of the celiac axis. Tumor emboli were found in distal aortic branches and most abdominal organs. Immunoperoxidase for Factor VIII and electron microscopy (EM) did not support an endothelial origin. EM showed myofibroblastic differentiation. Review of the literature yields an array of diagnostic histologic terms for these tumors, hampering case comparison. The literature does suggest, however, that the clinical presentation of these rare neoplasms correlates nicely with the location and gross morphology of the lesion. We therefore propose a clinicopathologic classification, categorizing the lesions as intimal (obstructive and nonobstructive) and mural. The former are typically pleomorphic sarcomas and are probably of myofibroblastic origin, whereas the latter are usually leiomyosarcomas or fibrosarcomas that probably originate in the media or adventitia.
Aorta, Abdominal↗
Mycobacterium xenopi and Mycobacterium kansasii in a hospital water supply.
A steady rise in the number of isolations of Mycobacterium xenopi from patients in a general hospital led to an examination of water taps. Most patients had been accommodated in a group of wards which had a common water supply. This organism was recovered from 35 of 69 outlets, mostly from hot and mixer taps in those wards. Mycobacterium kansasii was also isolated from 14, mostly cold and mixer taps. Ten strains of Myco. xenopi were recovered from 131 taps sampled at 10 other locations. We conclude that colonization of water supplies by mycobacteria is a likely source of contamination of clinical specimens.
England↗