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Biomedical subjects

N K Day

Publications and source records attributed to N K Day.

At least 19 recordsLinked to original sources
PubMed2001

Involvement of a herbimycin A-sensitive protein tyrosine kinase in extracellular action of HIV-1 Nef.

Extracellular Nef which has been implicated in disease progression and development of AIDS induces IL-10, a potent immunosuppressive cytokine, in vitro. The present study was designed to examine whether the action of extracellular Nef is a protein tyrosine kinase (PTK)-dependent event. Anti-phosphotyrosine immunoblotting reveals that recombinant HIV-1 Nef induces rapid tyrosyl phosphorylation of several cellular proteins in human peripheral blood mononuclear cells. Pre-treatment of cells with herbimycin A, but not with genistein, significantly abolishes the Nef-induced tyrosine phosphorylation of cellular proteins. Furthermore, ELISA and RNase protection assays show that herbimycin A significantly blocks Nef-induced production of IL-10 at both the protein and the mRNA level. Genistein and aminogenistein have a much less blocking effect on the ability of Nef to induce IL-10. These results provide evidence for the involvement of a herbimycin A-sensitive PTK in the signal transduction pathway for exogenous HIV-1 Nef.

Benzoquinones↗
PubMed2001

Immunologic reconstitution following bone marrow transplantation for X-linked hyper IgM syndrome.

X-linked hyper IgM syndrome (XHIM), caused by mutations of the CD40 ligand (CD40L) gene, is characterized by recurrent bacterial and opportunistic infections, an increased incidence of autoimmunity and malignancies, and immunodeficiency due to abnormal T/B cell interaction. Because of poor long-term prognosis, bone marrow transplantation (BMT) has been proposed as an alternative treatment. An 8-month-old boy with XHIM and a splice site mutation of CD40L underwent BMT using a fully matched sibling donor. Markers of engraftment and immunologic reconstitution were measured serially. After BMT, activated T cells expressed functional CD40L, and genomic DNA obtained from circulating white cells contained predominantly wild-type CD40L sequences. Serum immunoglobulin levels including IgE and antibody responses to recall antigens normalized, and immunization with the T-cell-dependent neoantigen, bacteriophage φX174, demonstrated amplification of the response and isotope switching. BMT provides a permanent cure for XHIM if a fully matched sibling donor is available and the procedure is performed before complications have occurred.

Bone Marrow Transplantation↗
PubMed2000

Monocytes are target cells for IL-10 induction by HIV-1 Nef protein.

IL-10 plays a pivotal role in the pathogenesis of several diseases and is elevated in sera of HIV-infected patients. Recently, we demonstrated that HIV Nef induces IL-10 mRNA expression as well as IL-10 production using PBMCs, H9 or U937 cells. This induction of IL-10 is inhibited by a calmodulin antagonist, W-7. In the present study, T or B lymphocytes or monocytes were isolated from PBMCs of healthy HIV-negative donors. Production of IL-10 and mRNA gene expression were analyzed on each isolated cell population after treatment with Nef or SEA for 3-24 h. The results show that Nef induces IL-10 production as well as mRNA expression significantly using monocytes but not with T or B lymphocytes. By contrast, SEA induced IL-10 production as well as mRNA expression using T lymphocytes but not with monocytes or B lymphocytes.

Avian Proteins↗
PubMed2000

DiGeorge anomaly: a comparative study of the clinical and immunologic characteristics of patients positive and negative by fluorescence in situ hybridization.

BACKGROUND: DiGeorge anomaly (DGA) is defined as a field defect characterized by dysmorphic facies, hypoparathyroidism, congenital heart defects, and a deficiency in cell-mediated immunity, usually associated with a microdeletion in chromosome 22q11.2. Data correlating clinical and genetic information, especially in terms of the extent of the immunodeficiency and infectious complications, are scant. OBJECTIVE: The goal of this study was to define the severity of the immunodeficiency and infectious illnesses in DGA patients with characteristic clinical and genetic findings and compare them with a similar group of patients without a microdeletion in chromosome 22q11.2. METHODS: A retrospective chart review of patients referred for evaluation of DGA to our immunology service from 1989 to 1995 was conducted. Clinical and immunologic data were collected from their initial evaluation. Patients meeting at least 3 of 4 of these criteria were considered to meet strict clinical diagnostic criteria for DGA, and the results of analysis for a microdeletion in chromosome 22q11.2 for each patient was noted. RESULTS: Sixteen of the 22 patients meeting strict clinical criteria for DGA were available for analysis for the microdeletion at chromosome 22q.11.2. Of these, 13 (81%) were positive by fluorescence in situ hybridization (FISH); 9 of 13 (69%) had low CD3 numbers, 6 of 10 assayed (60%) had low thymulin levels; 10 of 13 (77%) had low CD4 numbers, and 10 of 12 (83%) had absent or small thymus glands. B cells were increased in 9 of 13 (69%) patients. Mitogen and antigen responses were normal in 6 of 7 (86%) patients tested. Eight of 13 (62%) had a history of increased frequency of infectious illnesses. All had recurrent respiratory infections, including sinusitis, otitis media, and pneumonia. Three of the 16 patients tested (19%) were FISH negative. Two of 3 (67%) had low CD3 and CD4 numbers. B cells were elevated in all patients. All had recurrent respiratory infections, low thymulin levels, and absent thymus glands. CONCLUSIONS: Contrary to traditional descriptions, this group of clinically and genetically defined patients with DGA had a predominantly mild cell-mediated immunodeficiency syndrome usually associated with infections characteristic of humoral immunodeficiencies. The patients who were FISH positive did not differ significantly from those that were FISH negative in terms of clinical and immunologic findings or infectious complications.

Child↗
PubMed2000

Age-associated increase of basal corticosterone levels decreases ED2high, NF-kappaBhigh activated macrophages.

The proportion of cells with a high density of ED2 (ED2high cells) in peritoneal cells from old rats was significantly lower than that from young rats. The expression of major histocompatibility complex class II (MHC class II) molecules, the antigen presentation, production of interleukin (IL)-1beta and IL-6, and nuclear factor-kappaB activity in ED2high cells were markedly higher than those in cells with a low density of ED2 (ED2low cells), although no significant difference was observed in the expression of MHC class II molecules and the antigen presentation between ED2high cells from young and old rats. Meanwhile, basal corticosterone concentration in serum and glucocorticoid (GC) receptor mRNA expression in peritoneal cells increased significantly in old rats. The proportion of ED2high cells was increased by adrenalectomy in young rats. Furthermore, nuclear translocation of GC receptor was observed in ED2low cells, whereas GC receptor was detected in cytoplasmic extracts from ED2high cells. These results suggest that the decrease in functional ED2high macrophages with age results in the age-associated decline of immune responses, which is regulated, in part, by the basal GC concentration.

Adrenalectomy↗
PubMed1998

Interleukin 12 deficiency associated with recurrent infections.

A 3-yr-old female patient exhibited interleukin 12 (IL-12) deficiency that was associated with recurrent episodes of pneumococcal pneumonia with sepsis and other infections in the absence of fevers. The patient's peripheral blood mononuclear cells (PBMCs) exhibited normal proliferative responses to antigens. Immune responses, including in vivo production of antibodies to diphtheria, tetanus, or pneumococcal antigens, were normal. Ig levels and B cell and T cell phenotypes were also normal. In contrast, IL-12 p70 heterodimer production was undetectable by using supernatants of the patient's stimulated PBMCs when compared with control cells treated similarly. Although present, interferon gamma (IFN-gamma) was reduced. The addition of recombinant IFN-gamma to control cells enhanced the production of IL-12 by up to sixfold. By contrast, IL-12 was undetectable in supernatants of the patient's cells in the presence of recombinant IFN-gamma. IL-12 p40 subunit mRNA by using the patient's PBMCs after stimulation with Staphylococcus aureus Cowan strain 1 or lipopolysaccharide was also undetectable by reverse transcription-PCR when compared with control cells. Production of IL-2, IL-6, tumor necrosis factor alpha, or IFN-gamma of the patient's PBMCs after appropriate stimulation was observed. This patient has either a defect in Staphylococcus aureus Cowan strain 1-lipopolysaccharide- or staphylococcal enterotoxin A-induced signaling pathways for the activation of IL-12 p40 gene expression, or an abnormality in the IL-12 p40 gene itself.

Adolescent↗
PubMed1998

An increase in basal glucocorticoid concentration with age induces suppressor macrophages with high-density Fc gamma RII/III.

Ageing is usually accompanied by a decline in immune and neuroendocrine functions. To elucidate the mechanisms underlying age-related immunosuppression, the functions and surface phenotypes of peritoneal cells in the monocyte/macrophage lineage from old mice were investigated. The role of glucocorticoids (GC) in the immunomodulation was also examined. Proliferative responses of spleen cells from control mice stimulated with concanavalin A (Con A) were significantly suppressed by adding peritoneal exudate cells from old mice. Flow cytometry analysis revealed that the proportion of MAC-1+ cells with a high density of type II or type III receptor for the Fc portion of IgG (Fc gamma RII/IIIbright cells) was increased markedly in the periotoneal exudate cells from old mice. The prominent suppressor activity for Con A responses of control spleen cells was found in the Fc gamma RII/IIIbright cells, whereas MAC-1+ cells with a low density of Fc gamma RII/III (Fc gamma RII/IIIdull cells) did not suppress the Con A responses. On the other hand, both the basal corticosterone concentrations in serum and the mRNA expression for GC receptor in peritoneal exudate cells increased significantly in old mice. Furthermore, the proportion of Fc gamma RII/IIIbright cells in peritoneal exudate cells from old mice was normalized on administration of the GC antagonist RU 38,486 (mifepristone). These results suggest that the increase in basal corticosterone concentrations in old mice induces the generation of Fc gamma RII/IIIbright suppressor cells, possibly leading to the immune-suppressive state.

Aging↗
PubMed1997

Prolactin, epidermal growth factor or transforming growth factor-alpha activate a mammary cell-specific enhancer in mouse mammary tumor virus-long terminal repeat.

Mammary specific expression of elevated levels of mouse mammary tumor virus (MMTV) contributes to mammary carcinogenesis. Mechanisms which regulate provirus expression have not been completely defined. Using a MMTV-long repeat terminal (MMTV-LRT) directed chloramphenicol-acetyltransferase (CAT) reporter gene system and a human breast cancer cell line T47D, we demonstrate that prolactin (PRL), epidermal growth factor (EGF), or transforming growth factor-alpha (TGF-alpha) act on a mammary cell-specific enhancer at the extreme 5' end of the MMTV-LTR involving sequences -1094 through -858. PRL and either EGF or TGF-alpha exert concerted roles in this activation of these sequences. In contrast, using a plasmid construct lacking this mammary cell-specific enhancer, EGF or TGF-alpha, but not PRL, act synergistically with progesterone to induce CAT activity, indicating that the action of PRL on regulatory elements of the MMTV-LTR is restricted to this mammary cell-specific enhancer involving sequences -1094 through -858. A mobility shift assay was used to demonstrate that PRL, EGF or TGF-alpha induce nuclear factors (MP4, MAF, and MGF) which bind directly to this mammary cell-specific enhancer element.

Binding Sites↗
PubMed1997

Interleukin 10 is induced by recombinant HIV-1 Nef protein involving the calcium/calmodulin-dependent phosphodiesterase signal transduction pathway.

HIV-1 Nef protein shares a significant homology with the immunosuppressive and highly conserved retroviral transmembrane protein p15E. In the present study, extracellular Nef protein is shown to induce interleukin (IL)-10 mRNA expression in human peripheral blood mononuclear cells as well as in cells of H9 T and U937 promonocytic human cell lines. Release of IL-10 protein into supernatants of peripheral blood mononuclear cells stimulated with Nef is dose-dependent. Expression of cytokines IL-2, IL-4, IL-5, IL-12 p40, IL-13, and interferon gamma is not affected by Nef stimulation. IL-10 protein production induced by Nef is inhibited by the calcium/calmodulin phosphodiesterase inhibitor W-7 but not by the protein kinase A inhibitor H-89 nor the protein kinase C inhibitors staurosporine and calphostin C. The calcium chelating agent EGTA also inhibits the IL-10 production induced by Nef, and this inhibition is reversed by the addition of calcium along with Nef. These findings indicate that extracellular Nef may contribute to the immunopathogenesis of HIV infection by inducing IL-10.

Cell Line↗
PubMed1997

Immunosuppressive retroviral peptides: immunopathological implications for immunosuppressive influences of retroviral infections.

Studies of the effects of retroviruses on the immune system, which date back through thirty years of investigations, are reviewed. In the earliest published studies in the 1960s, it was demonstrated that mice infected with oncogenic viruses were immunosuppressed. Since then, numerous articles have been published describing profound immunodeficiencies observed in vivo in humans infected with human immunodeficiency virus and in animals such as cats infected with the feline immunodeficiency virus. In vitro investigations have shown that inactivated retroviruses or transmembrane envelope protein p15E as well as a synthetic 17-amino acid peptide (CKS-17) impressively conserved within the transmembrane envelope protein of several animal or human retroviruses are highly immunosuppressive. More recently, dysfunction of cytokines produced by CKS-17 at both a cellular and molecular level have been found to mimic influences observed in vivo in patients infected with the human immunodeficiency virus. CKS-17 has also been shown to induce cAMP in vitro. The significance of these observations to understanding the immunological disturbances observed in malignancy, cytokine biosynthesis, and modulations of immune functions through cAMP is discussed.

Amino Acid Sequence↗
PubMed1997

HIV-1 recombinant gp41 induces IL-10 expression and production in peripheral blood monocytes but not in T-lymphocytes.

The effects of recombinant gp41 (rgp41) protein of the human immunodeficiency virus type 1 (HIV-1) on interleukin 10 (IL-10) expression and production using human peripheral blood mononuclear cells was investigated. Expression of IL-10 mRNA was demonstrated within 3 h of cell exposure to endotoxin-free rgp41 by RT-PCR and Northern blot analyses in a time- and dose-dependent manner. IL-10 protein was detected in the supernatants of peripheral blood mononuclear cells following stimulation with rgp41 also in a dose dependent manner. Fractionation of peripheral blood mononuclear cells showed that purified monocytes but not purified T-lymphocytes induced expression of IL-10 mRNA by rgp41. Recombinant HIV-1 gp120 exhibits similar influences on the induction of IL-10. These results indicate that both of these components of envelope proteins may play an important role in HIV related immunomodulation by influencing regulatory functions of monocytes and macrophages.

Chemical Fractionation↗
PubMed1996

Diagnosis of pediatric human immunodeficiency virus infection by means of a commercially available polymerase chain reaction gene amplification.

OBJECTIVE: To assess the sensitivity and specificity of polymerase chain reaction (PCR) in infants and children at risk for human immunodeficiency virus (HIV) infection. DESIGN: A prospective, blinded study of 286 HIV-seropositive infants and children. Infection was diagnosed by antibody detection after 18 months of age, two positive direct tests (p24 antigen and HIV culture), or the presence of an illness that defines the acquired immunodeficiency syndrome. SETTING: University of South Florida and All Children's Hospital, St Petersburg, inpatient and outpatient centers. PARTICIPANTS: Two hundred eighty-six infants and children seropositive for HIV who were examined between July 1988 and September 1992. MAIN OUTCOME MEASURES: Sensitivity, specificity, and predictive values of a commercially available PCR test. RESULTS: Five hundred sixty-seven PCR tests were performed on samples from 286 seropositive subjects followed up for a minimum of 16 months. Of the subjects, 105 were confirmed to be infected and 181 uninfected. Overall, 96 of 105 initial PCRs in infected subjects were positive (sensitivity, 91.4%; positive predictive value, 99%). If samples obtained during the first week of life are excluded, 95 to 100 samples were positive (sensitivity, 95%). Of 181 initial PCR tests from seropositive subjects who seroreverted, 180 were negative (specificity, 99.4%,; negative predictive value, 95.2%). The predictive value of a positive test was 90.9% and that of a negative test was 93.1% in the first month of life. All 145 initial samples obtained between 5 weeks and 12 months of age correctly predicted infection status (positive predictive value, 100%). CONCLUSIONS: Gene amplification by means of a commercially available PCR is useful in the diagnosis of HIV infection for infants born to seropositive mothers. Between day 7 through 1 year of age, HIV infection is accurately diagnosed by the PCR assay.

Age Factors↗
PubMed1996

Normalization of cellular zinc levels in patients with Downs syndrome does not always correct low thymulin levels.

Retrospective analysis of five Down's syndrome (DS) patients who presented with recurrent infection revealed that all had initial low thymulin levels. Three patients had low cellular zinc levels that normalized after zinc replacement. Contrary to previous studies, thymulin levels were persistently low in four of five DS patients despite maintaining or achieving normal cellular zinc levels. A primary thymic epithelial defect may be responsible for the persistent thymulin deficiency in DS patients.

Adult↗
PubMed1996

Selective gamma-chain T-cell receptor gene rearrangements in a patient with Omenn's syndrome: absence of V-II subgroup (V gamma 9) transcripts.

Only gamma-chain T-cell receptor transcripts utilizing V-1 subgroup gene segments were found in peripheral blood lymphocytes from a patient with Omenn's syndrome. gamma-Chain T-cell receptor transcripts utilizing the V gamma 9 (V-II subgroup) gene segment were absent in peripheral blood lymphocytes from this patient. V gamma 9 J gamma 1.2 C gamma 1 rearrangements are those primarily found in peripheral blood lymphocytes (70 to 85%) from normal donors.

Eosinophilia↗
PubMed1996

Relationship between tumor necrosis factor alpha and feline immunodeficiency virus expressions.

The presence of feline immunodeficiency virus (FIV) proviral DNA, expression of FIV p26 core protein, and production of tumor necrosis factor alpha (TNF-alpha) were assessed in sequential biopsies of spleen and lymph node sections, of mononuclear cells of the peripheral blood, and of the serum of specific-pathogen-free cats during the acute phase of FIV infection. A temporal relationship between TNF-alpha production and FIV p26 expression was noted. Two months following FIV infection, and preceding the detection of FIV viremia, levels of TNF-alpha in serum increased significantly (P = 0.04), and they remained elevated during FIV viremia in the third month postinfection. Immunoprecipitates representing expression of TNF-alpha and of FIV p26 were localized in common foci of lymph nodes of FIV-infected cats during this period of active viremia. With the advent of anti-FIV antibodies, circulating levels of TNF-alpha and p26 antigen and expression of TNF-alpha and p26 in the lymph nodes decreased during the fifth month postinfection, and p26 production became undetectable. With clearance of viremia, burden of proviral DNA in peripheral blood mononuclear cells became reduced (P = 0.041), with provirus remaining integrated principally within lymph nodes (P = 0.046). During aviremia, p26 expression was undetectable in any tissue but remained inducible in vitro. During acute FIV infection, TNF-alpha production and p26 expression are intimately linked.

Animals↗