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Biomedical subjects

D T Purtilo

Publications and source records attributed to D T Purtilo.

At least 55 records · Page 3Linked to original sources

Current concepts of immunodeficiency disorders.

The technical developments in immunology and molecular biology during the 1970s, the growing use of organ transplantation during this period, and the abrupt emergence of AIDS in 1981 made the 1980s a fruitful period for investigating basic mechanisms of immunodeficiency. Now armed with powerful molecular biologic techniques, such as the polymerase chain reaction, we can identify specific genes and viruses that result in immunodeficiency. Complications associated with immunodeficiency, such as malignant lymphoma and opportunistic infections, have provided pathologists with opportunities for investigating the pathogenesis of this and other related diseases. An impetus for developing new diagnostic immunologic and molecular biological methods will be sustained during the 1990s. Pathologists working in the 21st century will be participants in the use of gene therapy for primary immunodeficiency diseases and for conquering AIDS. Perhaps with the declining threat of thermonuclear annihilation, we will tackle the largest cause of acquired immune deficiency, protein-calorie malnutrition.

Adult↗

Chromosome deletion of Xq25 in an individual with X-linked lymphoproliferative disease.

High resolution chromosome analysis was done on lymphoblastoid cell lines, established during the past decade from affected males with X-linked lymphoproliferative disease (XLP) or from obligate female carriers, from 14 families. One cell line, from a male with XLP, has a partial deletion of band Xq25. The constitutional nature of the deletion is confirmed in chromosome studies of peripheral blood from the affected individual and represents the first such structural defect to be described in this disorder. Cell lines from the remaining 13 families do not have cytogenetically detectable deletions. This observation will facilitate precise localization, cloning and sequencing of the gene causing XLP.

Chromosome Banding↗

Interferon-gamma in a family with X-linked lymphoproliferative syndrome with acute Epstein-Barr virus infection.

A 20-month-old male with fulminant infectious mononucleosis and the X-linked lymphoproliferative syndrome (XLP) was studied. Epstein-Barr virus (EBV)-determined nuclear antigen (EBNA) and EBV DNA were detected in various tissues. Despite a combined treatment with acyclovir, immunoglobulin, and methylprednisolone, the patient deteriorated rapidly. Following treatment with recombinant interferon-gamma (IFN-gamma), defervescence occurred and circulating EBNA-positive cells markedly decreased. IFN-gamma prior to treatment ranged from 10.8 to 24.5 U/ml in the patient's serum and increased linearly post exogenous IFN-gamma treatment. His natural killer (NK)-cell activity remained in the normal range throughout his illness but autologous EBV-infected cells were not killed in vitro by his peripheral blood lymphocytes (PBL). These results suggest that patients with the fatal infectious mononucleosis phenotype of XLP may produce endogenous IFN-gamma. Defective cytotoxic T cells against EBV-infected cells seem to be responsible for the fulminant infectious mononucleosis in this patient.

Acute Disease↗

Augmentation of natural cytotoxicity by leucine enkephalin in cultured peripheral blood mononuclear cells from patients infected with human immunodeficiency virus.

Natural Killer (NK) activity of lymphocytes from acquired immunodeficiency syndrome (AIDS) patients is frequently below normal and declines as disease progresses. We studied the potential of leucine enkephalin (leu-enkephalin) to restore this immune parameter by incubating nylon wool nonadherent mononuclear cells from 14 patients in the presence or absence of leu-enkephalin, and measuring NK cytolysis in a standard 51Cr release assay. The NK activity of human immunodeficiency virus antibody positive (HIV+) individuals with some remaining NK lytic ability was significantly augmented by leu-enkephalin concentrations of 10(-10) and 10(-8) M (n = 7). HIV+ patients with no measurable basal level of NK activity (n = 3) were not responsive to stimulation with leu-enkephalin. Human immunodeficiency virus antibody negative (HIV-) individuals (n = 4) responded in a pattern similar to normals. In addition, naloxone, an antagonist of alkaloid and peptide opiates including leu-enkephalin, displayed the properties of an antagonist/agonist, reflecting the immunoregulatory capacities of the endogenous opiate system.

Acquired Immunodeficiency Syndrome↗

Enzyme-linked immunosorbent assay (ELISA) for detecting antibodies in sera of patients with adenovirus infection.

The 41 distinct antigenic types of adenoviruses (Ads) are responsible for a broad spectrum of diseases in humans. We have developed an enzyme-linked immunosorbent assay (ELISA) using adenovirus (Ad) infected MRC-5 cells for detecting IgG and IgM antibodies to Ads. Using the ELISA, we detected IgG antibodies in 100% (20/20) of sera from normal adults (geometric mean titer, GMT = 1840.8, range = 40-20,480) and IgM antibodies in 3 of 20 sera (15%) with a GMT of 25.1. Our indirect immunofluorescence (IF) technique also detected IgG antibodies in 100% of these sera (GMT = 248.3, range = 40-5,120) and IgM antibodies in the 3 samples reactive in ELISA (GMT = 20.0, range = less than 5-40). In contrast, the complement fixation (CF) test detected antibodies to Ads in only 65% (13/20) of these sera (GMT = 10.9, range = less than 4-32). Moreover, IgG and IgM responses could not be distinguished using CF. Thus the sensitivity of these three techniques is greatest for ELISA. Additionally, a study of sequential sera from 3 patients with acute Ad infection disclosed seroconversion using all three methods. Both the ELISA and IF techniques permit the detection of transition from IgM to IgG, whereas CF only detects conversion from seronegativity to seropositivity. Finally, preliminary data suggest that the IgM response as measured by ELISA is specific for subgroups or types of Ad. This newly devised ELISA may be useful for detecting Ad infections.

Adenoviridae Infections↗

Detection of X-linked lymphoproliferative disease using molecular and immunovirologic markers.

PURPOSE, PATIENTS, AND METHODS: Detection of males affected with the X-linked lymphoproliferative disease (XLP) was sought using immunovirologic and molecular genetic linkage techniques. The study population consisted of 20 males in six families with XLP. RESULTS: Concordance for detection of affected males was 100% when linkage analysis using DXS42 and DXS37 DNA probes and antibody responses to challenge with bacteriophage phi X174 were both determined. Most affected males showing IgG subclass immune deficiency could not produce antibodies to Epstein-Barr virus nuclear antigen and had deficient responses to challenge with bacteriophage phi X174. CONCLUSION: Use of only one of the techniques described can fail to lead to the diagnosis of XLP, because problems can prevail with each individual determination.

Adolescent↗

Human herpesvirus 6 infection and Kawasaki disease.

Eighteen of a total of 22 serum specimens (81.8%) from patients with Kawasaki disease were positive for immunoglobulin G or M antibodies to human herpesvirus 6, whereas 10 of 16 age- and sex-matched healthy controls (62.5%) were seropositive. Additionally, increased geometric mean antibody titers of immunoglobulin G were shown in these patients. These results suggest that the status of human herpesvirus 6 infection may be a reflection of the immunologic alterations that are associated with Kawasaki disease.

Antibodies, Viral↗

Epstein-Barr virus-related lymphoproliferative disorders following bone marrow transplantation: an immunologic and genotypic analysis.

Four patients from 1.5 to 18 yr of age who had received partially matched T-cell-depleted bone marrow transplants for acute leukemia succumbed to a widespread lymphoproliferative disorder (LPD) at 56 to 147 days after transplant. Premortem diagnosis of LPD was suggested in two because plasmacytoid cells were observed in the blood and bone marrow, and in the cerebrospinal fluid of one of these patients. Serum clonal immunoglobulins (Igs) were also demonstrated in these two patients premortem, while the other two had clonal Igs in serum obtained at autopsy. Autopsies revealed a plasmacytoid infiltrate or immunoblastic lymphoma involving lymph nodes, spleen, liver, lungs, gastrointestinal tract, and kidneys. Immunoglobulin gene rearrangement studies performed in three revealed B-cell clonality. Both immunohistochemical and DNA gene rearrangement studies were useful in differentiating the LPD from the pretransplant leukemia. Epstein-Barr virus (EBV) genome was found in the tissues of the three patients studied. The diagnosis of EBV-induced LPD must be considered in bone marrow transplant patients who deteriorate and who exhibit serum clonal Igs or prominent plasmacytoid cells in laboratory specimens.

Adolescent↗

Association of DNA content and proliferative activity with clinical outcome in patients with diffuse mixed cell and large cell non-Hodgkin's lymphoma.

Formalin-fixed and paraffin-embedded lymph node biopsy specimens from 52 untreated patients with newly diagnosed diffuse large cell (n = 48) or mixed cell (n = 4) non-Hodgkin's lymphoma (NHL) were analyzed for DNA content and proliferative activity (PA) by flow cytometry. The results obtained by flow cytometry were compared with the results of cytogenetic studies performed on 28 of the specimens. The median age of the patients was 65 years (range, 15-84 years) and the male to female ratio was 3 to 2. All patients were uniformly staged and uniformly treated with cyclophosphamide, doxorubicin, procarbazine, bleomycin, vincristine, and prednisone. The flow cytometric results were compared statistically by univariate analysis with the rate and duration of complete remission and survival. Tumors with low PA (greater than or equal to 80% of cells in G0/G1 phase) were found in 65% of the patients; 74% of those with low PA versus only 44% of those with high PA achieved an initial complete remission (P less than 0.02). DNA aneuploidy was detected in tumors of 56% of the patients and was associated with a significantly longer duration of complete remission (P less than 0.01). Both low PA and aneuploidy independently predicted longer survival. The predicted 2-year actuarial survival for patients with tumors with low PA was 68% versus 10% for those with high PA (P less than 0.01). Similarly, the 2-year survival of patients with aneuploid tumors was 60% versus 36% for those with diploid tumors (P less than 0.01). The combination of PA and DNA content categorized the patients into four groups with decreasing 2-year survivals: low PA/aneuploid (n = 20), 77%; low PA/diploid (n = 14), 57%; high PA/aneuploid (n = 9), 32%; high PA/diploid (n = 9), 0%. The flow cytometric results correlated well with those of the cytogenetic studies. We conclude that low PA and DNA aneuploidy, both separately and in combination, predict a favorable clinical outcome for patients with diffuse mixed cell and large cell NHL.

Adolescent↗

Correlation of secondary cytogenetic abnormalities with histologic appearance in non-Hodgkin's lymphomas bearing t(14;18)(q32;q21).

Successful cytogenetic studies were performed on 69 biopsies from 64 patients with non-Hodgkin's lymphoma bearing a t(14;18)(q32;q21) translocation. This translocation appears to be a primary abnormality associated with the development of certain B-cell non-Hodgkin's lymphomas. We correlated the occurrence of secondary abnormalities, in addition to the t(14;18)(q32;q21), with histologic subtype to test the hypothesis that secondary abnormalities correlate with more aggressive histologic appearance. A large number of secondary abnormalities were identified, the most frequent being additional copies of chromosomes 7 (30%), 12 (22%), 18 (22%), 20 (16%), or 21 (14%), deletion of a portion of the long arm of chromosome 6 (17%), and either an additional chromosome 17 or an isochromosome for the long arm of chromosome 17 (13%). An extra chromosome 7 was highly associated with a diffuse histologic pattern; it was present in 52% of patients with a diffuse pattern and in only 15% of those with a follicular pattern (P = .002). A weaker association with a diffuse growth pattern was found for the addition of chromosome 17 or an i(17q); it was found in 24% of patients with a diffuse pattern and only 5% of those with a follicular pattern (P = .05). No other significant correlations between secondary chromosome abnormalities and histologic subtype were identified. Although the explanation for this association is not clear, it appears that patients with B-cell non-Hodgkin's lymphomas bearing the t(14;18)(q32;q21) translocation which also have an additional chromosome 7 are likely to exhibit a diffuse growth pattern.

B-Lymphocytes↗

Epstein-Barr virus and human diseases: recent advances in diagnosis.

Since the discovery of Epstein-Barr virus (EBV) from a cultured Burkitt's lymphoma cell line in 1964, the virus has been associated with Burkitt's lymphoma, nasopharyngeal carcinoma, and infectious mononucleosis. During the recent decade, EBV has been etiologically implicated in a broad spectrum of human diseases. The precise role of this virus in these diseases is not well understood, but clearly, defective immunosurveillance against the virus may permit an uncontrolled proliferation of EBV-infected cells. As a result, a growing number of cases of EBV-associated B-cell proliferative diseases or lymphoma have been noted in patients with primary and acquired immunodeficiencies. These lymphoproliferative diseases and others, such as chronic mononucleosis syndrome, are leading to new areas of investigation which are providing information regarding the pathogenetic mechanisms of EBV-induced diseases. The early accurate diagnosis of EBV infection can be achieved by performing EBV-specific serology, detecting for EBV-determined nuclear antigen in tissues, establishing spontaneous lymphoid cell lines, and using molecular hybridization techniques for demonstrating the presence of viral genome in affected lesions.

Burkitt Lymphoma↗

Adenovirus type-2 in a patient with lethal hemorrhagic colonic ulcers and chronic active Epstein-Barr virus infection.

An adenovirus type-2 was isolated from peripheral blood lymphocytes and throat washings from a patient with severe chronic active Epstein-Barr virus infection. Despite the Epstein-Barr virus reactivation, attempts to establish spontaneous lymphoblastoid cell lines from peripheral blood lymphocytes and to immortalize cord lymphocytes with throat washings were unsuccessful due to a marked cytopathic effect. The supernatants from the cultures induced cytopathic effect in cultured cord lymphocytes, MRC-5 cells, A-549 cells, or Vero cells. Virus particles with adenovirus morphology were seen by electron microscopy. Using type-specific antisera, the isolate was identified as adenovirus type-2. In addition, both Epstein-Barr virus and adenovirus type-2 genomes were seen in the colonic tissues and spleen. These results suggest that the combination of Epstein-Barr virus and adenovirus type-2 may be etiologic agents in the development of chronic active Epstein-Barr virus infection in this patient.

Adenoviridae Infections↗

Epstein-Barr virus--associated lymphoproliferative lesions.

From this review of the natural history of EBV infection in humans, it is clear that the virus has evolved to a symbiotic state with humans. Once individuals are infected, EBV establishes a permanent infection that is maintained at a low level by replication of the virus in the oropharyngeal region with subsequent seeding of circulating B-lymphocytes. Individuals with normal immune systems are able to control the pronounced proliferative potential of EBV-infected cells and thus prevent the emergence of lymphoproliferations. Disease states result when the immune system is altered by other infections, developmental conditions, immunosuppressive agents, or debilitating circumstances such as cancer or starvation. In some cases, localized lymphoproliferations resembling large-cell non-Hodgkin's lymphomas can result that are monoclonal by immunoglobulin gene rearrangement studies. Remarkably, many of the localized masses will regress if the immunosuppressive agents or condition can be ameliorated in these individuals. In patients with hereditary immune deficiencies, these localized masses can progress to a fatal disease without further cytogenetic events. Burkitt's lymphoma, which is associated with EBV infection, appears to result when EBV-driven lymphoproliferations undergo cytogenetic translocations involving predominantly chromosome 8. Most of the conditions that are associated with EBV can be diagnosed by accurate application of EBV-serology, examination of peripheral blood films, a careful history and physical examination, and, in some cases, more sophisticated techniques such as the establishment of lymphoblastoid cell lines, EBNA staining, DNA probes, and pedigree analysis.

Antibodies, Viral↗

Soluble interleukin 2 receptors in patients with malignant lymphoma.

We performed a retrospective analysis of serum interleukin 2 receptor (IL-2R) levels in a group of 35 patients with malignant lymphoma (ML; 13 T cell and 22 B cell) using a new enzyme immunoassay. Our objectives were to determine if elevated levels of soluble IL-2R occur in patients with active ML, whether serum IL-2R levels are prognostic, and whether prospective studies are warranted. Our preliminary data indicate that serum IL-2R levels correlate with disease activity and size of tumor, but not with grade or stage of the tumor. Five-year actuarial survival was 20% for patients with IL-2R levels greater than 1000 U/mL at any time during their course and 86% for patients who did not exceed that threshold. Furthermore, patients with IL-2R levels lower than 1000 U/mL were more likely to achieve a complete remission. Serum lactate dehydrogenase and uric acid levels did not show significant correlation with disease activity or prognosis. We conclude that serum IL-2R levels may have clinical and prognostic significance in patients with ML and that prospective studies are indicated.

Antibodies, Viral↗

Soluble interleukin 2 receptors in sera of Japanese patients with adult T cell leukemia mark activity of disease.

Serum concentrations of soluble interleukin 2 receptors (sIL 2R) were measured by an enzyme-linked immunosorbent assay (ELISA) in 30 patients with adult T cell leukemia (ATL), in 9 patients with other hematopoietic malignancies, and in 17 asymptomatic individuals seropositive for human T cell leukemia virus type I (HTLV-I). Sixty HTLV-I seronegative, age-matched controls showed a normal range of form 63.2 to 480.8 U/mL. All asymptomatic carriers of HTLV-I had sIL 2R in their sera within the normal range. sIL 2R in sera was not related to the anti-HTLV-I antibody titer. Eleven patients with acute ATL, a clinical phenotype with median survival rate of 4.4 months, had markedly elevated sIL 2R (11,100 to 99,000 U/mL), but eight patients with smoldering ATL had low sIL 2R values (less than 480.8 U/mL) comparable to controls. Eleven patients with chronic ATL had intermediate elevated levels of sIL 2R (480.8 to 37,300.0 U/mL). Serum levels of sIL 2R correlated with the number of ATL cells (r = 0.812) and CD25-positive cells (r = 0.725) circulating in the peripheral blood. Longitudinal studies performed in four patients with ATL showed significant correlation between serum concentration of sIL 2R and activity of the malignancy. These findings suggest that the level of sIL 2R in serum indicated tumor load and, possibly, prognosis.

Adult↗

Bone marrow findings in immunodeficiency syndromes.

We have summarized the findings in selected congenital and acquired immunodeficiency disorders that frequently display blood and bone marrow abnormalities. Many other immunodeficiency syndromes were not discussed, as bone marrow abnormalities are infrequently seen in them. Examination of the bone marrow is not usually critical for diagnosing most immunodeficiency disorders. However, it may be diagnostic in the evaluation of fever of unknown origin, to rule out other causes of peripheral cytopenias, and to screen for virus-associated malignancies frequently seen in immune-deficient patients (for example, B cell lymphoma, Kaposi's sarcoma). Examination of the bone marrow is critical in the diagnosis of the virus-associated hemophagocytic syndrome and is often important in evaluating patients with the acquired immunodeficiency syndromes who display hematologic disturbances, such as those being treated with AZT, or those who might have opportunistic infections or malignant neoplasms.

Bone Marrow↗

Initial and subsequent cytogenetic studies in malignant lymphoma.

Two or more cytogenetic studies of lymph node biopsy specimens from 12 patients with non-Hodgkin's malignant lymphoma were performed. Each of the 12 patients had at least one clonal structural chromosome abnormality in their initial biopsy specimen. A comparison of the chromosome abnormalities in the initial lymph node biopsy specimens with those in subsequent biopsy specimens was made. Structural and numerical chromosome changes were not always accompanied by a change in the histologic type of the lymphoma. Similarly, a change in histologic type was not always associated with the appearance of new structural or numerical abnormalities. Serial cytogenetic studies from additional patients will further our understanding relevant to which chromosome changes are associated with lymphomagenesis and which are secondary or more of an evolutionary nature.

Adult↗