Immunohistologic staining of cytotoxic T and NK cells in formalin-fixed paraffin-embedded tissue using microwave TIA-1 antigen retrieval.
Explore the source record for details and available documents.
Formaldehyde↗
Biomedical subjects
D Myerson
Publications and source records attributed to D Myerson.
Early detection and treatment of cytomegalovirus infections in marrow transplant patients: methodological aspects and implications for therapeutic interventions.
Major advances have been made in the early detection of CMV infection after marrow transplant by the introduction of the polymerase chain reaction (PCR) and the CMV antigenemia assay. Numerous studies have established clinical correlations of these techniques but there is a need for technical standardization as assay sensitivity may vary considerably with different modifications. Current prevention strategies for CMV disease after marrow transplantation, including options on how the antigenemia assay and PCR assays may be used in early treatment strategies, are discussed.
Acyclovir↗
Cytomegalovirus infection of the tongue following marrow transplantation.
Tongue ulcerations in seven patients who had undergone allogeneic BMT for hematologic or lymphoid malignancies were examined for the presence of CMV. The clinical presentation of these tongue lesions was nonspecific and showed ulcerations similar to those associated with severe preparative conditioning regimen-related mucositis, HSV infection and oral acute GVHD. Tissue biopsies were studied by routine histology, immunocytochemistry for CMV and HSV antigens, in situ hybridization for CMV nucleic acid and standard as well as centrifugation viral cultures. Five of the 7 patients had lesions which were positive for CMV. While CMV oral lesions are known to occur in patients with the acquired immune deficiency syndrome (AIDS), these findings will improve our ability to recognize and diagnose tongue lesions in BMT patients and indicate that CMV should be considered in the differential diagnosis for similar ulcerations in other immunocompromised patients.
Adult↗
Diagnosis of cytomegalovirus pneumonia by the polymerase chain reaction with archived frozen lung tissue and bronchoalveolar lavage fluid.
The authors developed a polymerase chain reaction (PCR) procedure to detect cytomegalovirus (CMV) in archived, frozen lung tissue and bronchoalveolar lavage (BAL) fluid preparations. The procedure incorporated an internal beta-globin control to assess the adequacy of the sample. Twenty-nine lung tissue and 96 archived BAL specimens from marrow transplant recipients were tested. One of the lung tissue specimens and 46 of the BAL specimens had insufficient tissue for PCR analysis because they did not show a beta-globin band. In lung tissue, the PCR had a sensitivity of 100% and specificity of 89% compared with conventional tube culture. In BAL specimens, the PCR had a sensitivity of 87% and specificity of 90% compared with conventional tube culture or centrifugation culture. Provided tissue was sufficient, the results suggest that the PCR can be used to diagnose CMV interstitial pneumonia effectively from frozen specimens of lung tissue or BAL fluid.
Base Sequence↗
Cytomegalovirus-induced hemorrhagic cystitis following bone marrow transplantation.
Cytomegalovirus (CMV) is a common cause of morbidity and mortality following bone marrow transplantation but has not been demonstrated to cause hemorrhagic cystitis in this setting. We describe a patient who developed gross hematuria and lower abdominal pain 47 days after bone marrow transplantation was performed. Subsequently, CMV was detected in bladder endothelial cells with use of monoclonal antibody staining that was specific for the virus. No other cause for the patient's hemorrhagic cystitis was discovered. The gross hematuria and pain persisted until the patient received intravenous ganciclovir. This case demonstrates that CMV-induced hemorrhagic cystitis can occur following bone marrow transplantation and may respond to antiviral therapy.
Adolescent↗
A c-kit ligand, recombinant human stem cell factor, mediates reversible expansion of multiple CD34+ colony-forming cell types in blood and marrow of baboons.
The ligand for the human c-kit, recombinant human stem cell factor (SCF), was administered to baboons at doses of 200, 100, 50, 25, and 10 micrograms/kg/d. SCF induced a dose-dependent expansion of hematopoietic colony-forming cells (CFC) of multiple types in both blood and marrow, including colony-forming unit (CFU) granulocyte-monocyte, burst-forming unit-erythroid, CFU-MIX, and high proliferative potential-CFC. These changes were associated with a dose-dependent leukocytosis, involving all leukocyte lineages, a reticulocytosis, and increases in marrow cellularity. At 200 micrograms/kg/d of SCF, CFC in blood were increased 10-fold to greater than 100-fold. This correlated with an increased frequency of CD34+ cells in blood. The frequency of CFC in blood approached that of marrow in some animals. These changes were reversed within 7 to 14 days of stopping SCF. The results of these studies suggest a role for the c-kit ligand in stimulating the expansion of multiple CFC types in blood and marrow for potential therapeutic purposes.
Animals↗
Cytomegalovirus and marrow function.
Infection with cytomegalovirus (CMV) continues to be one of the most common complications following allogeneic bone marrow transplantation. A proportion of patients with CMV infection also experience neutropenia. To investigate the possible role of CMV in the suppression of hematopoiesis, we have examined the effect of CMV on the growth of isolated myeloid progenitors and on the production of myeloid cells in the long-term bone marrow culture (LTMC) system. In these studies, various isolates of CMV were added either directly to cultures of progenitors or to LTMC established from normal CMV-seronegative donors. In the first system, myelosuppression is manifested by a reduction in the number of colonies that grow. In the second system, myelosuppression is manifested by a reduction in the number of myeloid cells produced and released into the culture supernatant. Analysis of the data observed indicated that myelosuppression could in some cases be attributed to direct infection of myeloid progenitors. In other cases stromal cells were infected. In the latter cases, myelosuppression was then caused by an alteration in cytokines produced by the stromal cells. These observations made in vitro raise the possibility that comparable mechanisms may be responsible for the myelosuppression observed with CMV infection in vivo. To pursue this possibility we proposed to detect the CMV genome in defined subpopulations of marrow cells isolated from infected patients. Given the technical restrictions imposed by the small sample size available from patient marrow aspirations, our initial attempts to develop on appropriate technique involved isolation of cells from CMV-seropositive normal bone marrow donors. Using the polymerase chain reaction we were able to amplify CMV DNA contained within marrow cells of some healthy CMV-seropositive marrow donors.
Bone Marrow↗
Recurrent genital infection in the guinea pig: differences between herpes simplex types 1 and 2.
Recurrence rates of genital infections are significantly higher for herpes simplex virus (HSV) type 2 than HSV type 1. Reasons for this difference are not known. In this report, multiple strains of HSV-1 and HSV-2 were evaluated in the guinea-pig model. HSV-2 strains showed significantly higher genital lesion recurrence than HSV-1, including HSV-1 McKrae strain which is highly recurrent in ocular infections. HSV-2 strains were also associated with more frequent asymptomatic vaginal virus shedding. Further study showed that HSV-1 strains replicated as well as HSV-2 in both the genital tract and the nervous system during acute infection. In addition, no difference was detected between HSV-1 and HSV-2 in nervous system latency. Thus, a number of possible explanations for the observed difference in genital herpes recurrence rates were examined and excluded.
Acute Disease↗
Automation of in situ hybridization.
The authors used automated DNA hybridization equipment, the Code-On (Instrumentation Laboratory, Lexington, MA), for more than 2 years to perform in situ hybridization in the clinical and research laboratory. For in situ hybridization for viral DNA in fixed, paraffin-embedded tissue, the Code-On produces results that are as sensitive as the manual method and with considerably greater ease. The procedure must be modified to fit the operating characteristics of the Code-On. The authors outline a procedure that emphasizes sensitivity, rather than speed. The automated procedure requires close technical attention, but the authors propose that it is considerably more efficient than the manual method. One technician can produce reliable results on as many as 60 slides a day. For in situ hybridization on cytogenetic preparations, the results are excellent, but the procedure is contorted and the probe use is increased. For these reasons the Code-On is not used for routine interphase cytogenetics. The Code-On is in routine use in the authors' pathology laboratory for performing in situ hybridization on formalin, B5, and Carnoy's fixed, paraffin-embedded specimens.
Automation↗
Evidence of cytotoxic T-cell destruction of epidermal cells in human graft-vs-host disease. Immunohistology with monoclonal antibody TIA-1.
A newly described mouse monoclonal IgG1 antibody, TIA-1, binds serine esterase-positive granule membranes of cytotoxic human T cells and is a candidate for an effector molecule involved in T-cell cytolytic mechanisms analogous to those of the perforin system. We performed immunohistologic studies on frozen human skin (n = 5) and lip (n = 21) sections as well as on control frozen sections of tonsils, purified cytolytic T (CD8) cells, and B cells using an indirect immunoperoxidase system. We found a strong association of TIA-1+ cells with CD8+ cells invading the epidermis in lip and skin lesions of graft-vs-host disease in human marrow allograft recipients, as well as a sharp geographic association of TIA+ lymphocytes with CD8+ regions in human tonsil sections. Double staining of CD8 and TIA-1 antigens with fluorescein isothiocyanate and Texas red confirmed that 80% to 90% of the CD8 cells were TIA-1+ in the epidermal infiltrates. Leu-7 activity (natural killer cell) was minimal and found in only three of 17 lip biopsy specimens. These data provide new evidence that direct cytolytic attack by donor T lymphocytes is the mechanism of epithelial target cell killing in human graft-vs-host disease.
Adolescent↗
Allogeneic marrow transplantation in patients positive for hepatitis B surface antigen.
Twenty patients who were positive for hepatitis B surface antigen (HBsAg) underwent allogeneic marrow transplant for malignancy or other underlying hematologic disease between 1975 and 1986. After transplant, one patient had serologic evidence of hepatitis B virus (HBV) reactivation whereas three patients had evidence of an immune response to HBV. Among four patients with serologic follow-up of 1 year or more, three remained positive for HBsAg and one became HBsAg negative. Six patients (30%) developed clinical evidence of venocclusive disease and seven patients (35%) developed acute graft-versus-host disease involving the liver, but the incidence of these complications was similar to that expected among patients who are not carriers of HBsAg. Three patients died with hepatorenal failure, but all three had venocclusive disease and the contribution of HBV infection to liver failure was unclear. Available liver specimens obtained at autopsy (six patients) or biopsy (two patients) all showed either HBsAg (one specimen) or hepatitis B core antigen (four specimens) or both (three specimens) by immunoperoxidase staining. Although HBV reactivation leading to hepatic failure has been reported among allogeneic marrow transplant recipients as well as other immunocompromised patients, we did not observe an increase in the incidence of severe liver disease after transplant among these 20 patients positive for HBsAg at the time of transplant, and do not consider positivity for HBsAg to be a contraindication to allogeneic marrow transplantation.
Adolescent↗
Comparative virologic studies of condylomata acuminata reveal a lack of dual infections with human papillomaviruses.
Condylomata acuminata are epithelial proliferations caused by infection of the anogenital squamous epithelium with human papillomavirus (HPV). DNA-DNA hybridization techniques and the extremely sensitive polymerase chain reaction (PCR) were used to analyze biopsies from patients with clinically diagnosed condyloma acuminatum for the presence of HPV DNA. PCR analyses using primers and oligonucleotide probes specific for the E6/E7 region of HPV-6, -11, or -16 showed that 31 (93.9%) of 33 tissue biopsies contained HPV DNA: 22 contained type 6 DNA, 6 contained type 11 DNA, and 3 contained type 16 DNA. Eleven biopsies positive by PCR were Southern hybridization-negative or were considered inadequate for Southern analysis. In all 11, the presence of HPV DNA was corroborated by the observation of histopathologic evidence suggestive of HPV infection or by in situ hybridization. No evidence of multiple infections with HPV-6 or -11 and HPV-16 was seen.
Adult↗
Producing single-stranded DNA probes with the Taq DNA polymerase: a high yield protocol.
We report an efficient procedure to synthesize either single- or double-stranded probes labeled with biotin-11-dUTP, biotin-21-dUTP or digoxigenin-11-dUTP. To produce the single-stranded probe, only a single primer is utilized in a Taq polymerase amplification of 55 cycles. A cytomegalovirus probe is presented. This procedure allows easy production of nonradioactively labeled pure single-stranded probes of any desired length and specificity.
Base Sequence↗
Endothelial changes in cutaneous graft-versus-host disease: a comparison between HLA matched and mismatched recipients of bone marrow transplantation.
Histological features of endothelial and vascular changes seen in cell mediated immune reactions were evaluated in skin biopsies from 44 HLA matched and mismatched recipients of bone marrow transplantation. The mismatched group (n = 24) was further subdivided into 'D-mismatched' (n = 14) and 'AB-mismatched' (n = 10). All patients had clinical grade III and histological grade II graft-versus-host disease (GVHD). Comparison groups consisted of skin biopsies from 10 non-GVHD cases. Histological evidence of endothelial damage including perivascular factor VIII related antigen deposition was assessed and compared between the groups. The results showed that all patients post-transplant had evidence of a non-specific vascular response consisting mainly of a perivascular lymphocytic infiltrate, perivascular oedema and factor VIII related antigen extravasation. However, a significant difference between matched and mismatched patients who developed GVHD was the more frequent observation of perivascular nuclear dust in the HLA mismatched group (p less than 0.01) suggesting a greater degree of endothelial cell damage in these patients.
Bone Marrow Transplantation↗
Human papillomavirus infection and anal cancer.
To study the association of human papillomavirus (HPV) infection with anal cancer, we examined tissue specimens from 126 patients with malignant lesions of the anal skin or mucosa. The patients were enrolled in a population-based, case-control study of ano-rectal cancer which is being conducted in the state of Washington and the Province of British Columbia. Histologic sections from formalin-fixed, paraffin-embedded tissues were tested for the presence of HPV DNA by in situ hybridization with biotin-labelled HPV 6, 11, 16, 18 and 31 DNA probes. HPV DNA sequences were found in tumor tissues from 24 of the 126 subjects (19.0%). When only squamous neoplasms are considered, 23 of 70 subjects (32.9%) had lesions which contained detectable HPV DNA. One HPV-positive patient had a cloacogenic carcinoma that contained regions of squamous differentiation and it was in these squamous cells that HPV DNA was localized. Of the 23 squamous lesions that harbored detectable HPV DNA, 8 contained HPV 6, 10 contained HPV 16, 1 contained HPV 18 and 4 contained an unclassified virus type(s). HPV DNA was found in tissues from 14 patients with carcinoma-in situ and 10 subjects with invasive carcinoma. These results demonstrate that some malignant tumors of the anus, in both men and women, are associated with HPV infection. We conclude that the anal squamous epithelium is another site where infection with the common genital tract HPVs may carry a risk of malignant transformation.
Adenocarcinoma↗
Presence of human papillomavirus deoxyribonucleic acid.
Explore the source record for details and available documents.
DNA, Viral↗
Direct detection of cytomegalovirus from bronchoalveolar lavage samples by using a rapid in situ DNA hybridization assay.
An in situ DNA hybridization assay was compared with centrifugation culture for rapid detection of cytomegalovirus (CMV) from bronchoalveolar lavage (BAL) samples. Eighty BAL samples were inoculated into both centrifugation culture and standard culture. Cytospin preparations of the BAL samples were studied in a 75-min in situ DNA hybridization assay using the PathoGene CMV kit (Enzo Biochem, Inc., New York, N.Y.). Of the 80 samples, 39 (49%) were positive for CMV; 37 of 39 (95%) were positive by centrifugation culture, 34 of 39 (87%) were positive in standard culture, 24 of 39 (62%) were positive by in situ hybridization, and 20 of 39 (56%) were positive by histologic and/or immunofluorescence techniques. The in situ hybridization assay detected 23 of the 37 samples positive in centrifugation culture, for a sensitivity of 62% and a specificity of 98%. We conclude that the in situ hybridization assay is a specific and more rapid test than centrifugation culture and standard culture for diagnosis of CMV pulmonary infection. For the clinical laboratory, however, current hybridization methods are not sufficiently sensitive to replace centrifugation culture for detection of CMV in BAL specimens.
Bronchoalveolar Lavage Fluid↗
Self-limited cytomegalovirus colitis in immunocompetent individuals.
We report 3 cases of isolated acute cytomegalovirus colitis in immunocompetent individuals. Each case was documented by the presence of cytomegalovirus inclusions in colonic biopsy specimens. All had a self-limited illness. Two of these cases occurred after anal intercourse and 1 case had no probable known cause. At follow-up from 2 to 6 yr later, none of the 3 patients has had any gastrointestinal symptoms. Additional biopsy specimens were obtained from 2 patients and no evidence of residual cytomegalovirus was seen by light microscopy or by in situ hybridization. Although cytomegalovirus colitis in immunosuppressed individuals is well recognized, these are the first reports of a nonfatal cytomegalovirus colitis in immunocompetent hosts.
Acute Disease↗